Methods for treating hypercholesterolemia

ABSTRACT

Disclosed herein are antisense compounds and methods for decreasing LDL-C in an individual having elevated LDL-C. Additionally disclosed are antisense compounds and methods for treating, preventing, or ameliorating hypercholesterolemia and/or atherosclerosis. Further disclosed are antisense compounds and methods for decreasing coronary heart disease risk. Such methods include administering to an individual in need of treatment an antisense compound targeted to a PCSK9 nucleic acid. The antisense compounds administered include gapmer antisense oligonucleotides.

REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No. 61/059,169, filed Jun. 5, 2008, and U.S. Provisional Application No. 61/058,707, filed Jun. 4, 2008, which are herein incorporated by reference in their entirety. This application is a continuation-in-part of U.S. application Ser. No. 12/516,457, filed Jan. 20, 2010, which is a National Stage application of International PCT Application No. PCT/US07/024369, now WO 2008/066776, filed Nov. 27, 2007, which claims the benefit of U.S. Provisional Application No. 60/988,074, filed Nov. 14, 2007, U.S. Provisional Application No. 60/986,286, filed Nov. 7, 2007, U.S. Provisional Application No. 60/912,892, filed Apr. 19, 2007, and U.S. Provisional Application No. 60/867,395, filed Nov. 27, 2006, which all are herein incorporated by reference in their entirety.

FIELD OF THE INVENTION

Methods are provided for lowering LDL-C levels in an individual having elevated LDL-C levels. Such methods are further useful to treat hypercholesterolemia, and to reduce the risk of coronary heart disease.

SEQUENCE LISTING

The Sequence Listing attached below is herein incorporated by reference in its entirety.

BACKGROUND

Atherosclerosis is a complex, polygenic disease of arterial degeneration that can lead to coronary heart disease (CHD). In Western societies, complications arising from atherosclerosis are the most common causes of death. Several risk factors for CHD have been well-established, and include elevated low density lipoprotein-cholesterol (LDL-C), low levels of high density lipoprotein cholesterol (HDL-C), cigarette smoking, hypertension, age, and family history of early CHD. Given the abundant clinical data and epidemiological studies indicating that lowering LDL-C is beneficial for the prevention of adverse coronary events, the primary target of pharmacological intervention in the treatment and prevention of CHD is lowered LDL-C.

In individuals with autosomal dominant hypercholesterolemia (ADH), elevated LDL-C levels have been linked to mutations in the genes encoding LDL-receptor (LDL-R), apolipoprotein B (apoB), or proprotein convertase subtilisin/kexin type 9 (PCSK9) (Abifadel et al., Nat. Genet., 2003, 34:154-156). The PCSK9 gene (also known as FH3; NARC1; NARC-1; and HCHOLA3) was mapped to Chromosome 1 at location 1p32.3. PCSK9 was identified as a third locus associated with ADH when gain-of-function mutations in PCSK9 were found to be linked to elevated LDL-C levels. ApoB-100 participates in the intracellular assembly and secretion of triglyceride-rich lipoproteins and is a ligand for the LDL-R. PCSK9 is proposed to reduce LDL-R expression levels in the liver. Reduced LDL-R expression results in reduced hepatic uptake of circulating apoB-100-containing lipoproteins, which in turn leads to elevated cholesterol.

Familial hypercholesterolemia (FH) is caused by hundreds of different mutations in the LDL-R, and is phenotypically characterized by elevated plasma LDL-C levels and deposits of LDL-C in tendons, skin and arteries, leading to premature cardiovascular disease. Homozygous and heterozygous mutations in the LDL-R are associated with FH. Likewise, heterozygous and homozygous mutations in the ligand-binding domain of PCSK9 are associated with the FH phenotype. Mutations in this gene have been associated with a third form of autosomal dominant FH.

In mice genetically deficient in PCSK9, a marked increase in LDL-R expression and increased plasma LDL clearance rate were observed. Conversely, overexpression in mice of wild-type PCSK9, or certain PCSK9 mutants, promotes decreased LDL-R expression and elevated LDL-C levels.

SUMMARY OF THE INVENTION

Provided herein are methods for the treatment of hypercholesterolemia and/or atherosclerosis, as well as methods for the reduction of elevated LDL-C levels and CHD risk. Methods for treating hypercholesterolemia comprise administering to an individual an antisense compound targeted to a PCSK9 nucleic acid. In such methods, an antisense compound targeted to a PCSK9 nucleic acid may be targeted to sequences as set forth in GENBANK® Accession No. NM_(—)174936.2, nucleotides 25475000 to 25504000 of GENBANK® Accession No. NT_(—)032977.8, or GENBANK® Accession No. AK124635.1.

Methods for treating and/or preventing atherosclerosis comprise administering to an individual an antisense compound targeted to a PCSK9 nucleic acid. Methods for reducing LDL-C levels comprise administering to an individual an antisense compound targeted to a PCSK9 nucleic acid. Methods for reducing CHD risk comprise administering to an individual an antisense compound targeted to a PCSK9 nucleic acid. Such methods may comprise the administration of a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid.

Methods are provided for reducing LDL-C levels in an individual having elevated LDL-C levels, comprising administering to the individual a therapeutically effective amount of an antisense oligonucleotide targeted to a PCSK9 nucleic acid, thereby reducing LDL-C levels. Also provided are methods for reducing LDL-C levels in an individual comprising selecting an individual having elevated LDL-C levels, and administering to the individual a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid, and additionally monitoring LDL-cholesterol levels. Further provided are methods for treating atherosclerosis in an individual, comprising selecting an individual diagnosed with atherosclerosis, administering to the individual a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid, and monitoring atherosclerosis. Also provided are methods for reducing coronary heart disease risk, comprising selecting an individual having elevated LDL-C levels and one or more additional indicators of coronary heart disease, administering to the individual a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid, and monitoring LDL-C levels. Additionally provided are methods for treating hypercholesterolemia, comprising administering to an individual diagnosed with hypercholesterolemia a therapeutically effective amount of an antisense oligonucleotide targeted to a PCSK9 nucleic acid, thereby reducing cholesterol levels.

Further provided is a use of an effective amount of one or more of the disclosed antisense compounds for treating atherosclerosis or hypercholesterolemia in an individual in need of such treatment. Also provided is a use of an effective amount of one or more of the disclosed antisense compounds for reducing LDL-C levels or reducing CHD risk in an individual in need of such treatment. Also provided is the use of one or more of the disclosed antisense compounds in the manufacture of a medicament for the treatment of atherosclerosis or hypercholesterolemia. Further provided is the use of one or more of the disclosed antisense compounds in the manufacture of a medicament for reducing LDL-C levels or for reducing CHD risk.

In any of the methods provided, a PCSK9 nucleic acid may be the sequence set forth in SEQ ID NO: 1. Thus, the antisense compound may be targeted to a PCSK9 nucleic acid as set forth in SEQ ID NO: 1.

Any of the methods provided herein may further comprise monitoring LDL-C levels.

An individual may be selected for administration of an antisense compound targeted to a PCSK9 nucleic acid when the individual exhibits an LDL-C level above 100 mg/dL, above 130 mg/dL, above 160 mg/dL, or above 190 mg/dL. Administration of an antisense compound targeted to a PCSK9 nucleic acid may result in an LDL-C level below 190 mg/dL, below 160 mg/dL, below 130 mg/dL, below 100 mg/dL, below 70 mg/dL, or below 50 mg/dL.

In any of the aforementioned methods, administration of the antisense compound may comprise parenteral administration. The parenteral administration may further comprise subcutaneous or intravenous administration.

In any of the methods provided herein, the antisense compound may have least 80%, at least 90%, or at least 95% complementarity to SEQ ID NO: 1. Alternatively, the antisense compound may have 100% complementarity to SEQ ID NO: 1.

The antisense compounds provided herein and employed in any of the described methods may be 8 to 80 subunits in length, 12 to 50 subunits in length, 12 to 30 subunits in length, 15 to 30 subunits in length, 18 to 24 subunits in length, 19 to 22 subunits in length, or 20 subunits in length. Further, the antisense compounds employed in any of the described methods may be antisense oligonucleotides 8 to 80 nucleotides in length, 12 to 50 nucleotides in length, 12 to 30 nucleotides in length 15 to 30 nucleotides in length, 18 to 24 nucleotides in length, 19 to 22 nucleotides in length, or 20 nucleotides in length.

In any of the methods provided, the antisense compound may be an antisense oligonucleotide. Moreover, the antisense oligonucleotide may be a gapmer antisense oligonucleotide. The gapmer antisense oligonucleotide may comprise a gap segment of ten 2′-deoxynucleotides positioned between wing segments of five 2′-MOE nucleotides. The gapmer antisense oligonucleotide may be a gap-widened antisense oligonucleotide, comprising a gap segment of fourteen 2′-deoxynucleotides positioned between wing segments of three 2′-MOE nucleotides.

In any of the methods provided, the antisense compounds may have at least one modified internucleoside linkage. Additionally, each internucleoside linkage may be a phosphorothioate internucleoside linkage. Each cytosine may be a 5-methyl cytosine.

Also provided herein are antisense compounds targeted to a PCSK9 nucleic acid. Further provided are antisense oligonucleotides targeted to a PCSK9 nucleic acid. The antisense compounds, including antisense oligonucleotides, may be targeted to PCSK9 nucleic acids, which include the sequences as set forth in GENBANK® Accession No. NM_(—)174936.2, nucleotides 25475000 to 25504000 of GENBANK® Accession No. NT_(—)032977.8, or GENBANK® Accession No. AK124635.1. The antisense compounds, including antisense oligonucleotides, may have at least 70%, at least 80%, at least 90%, or at least 95% complementarity to a PCSK9 nucleic acid. The antisense compounds, including antisense oligonucleotides, may have 99% complementarity to a PCSK9 nucleic acid. The antisense compounds, including antisense oligonucleotides, may have 100% complementarity to a PCSK9 nucleic acid. For any of the antisense compounds provided, including antisense oligonucleotides, the PCSK9 nucleic acid may be the sequence set forth in SEQ ID NO: 1.

Antisense compounds targeted to a PCSK9 nucleic acid may be 8 to 80 subunits in length, 12 to 50 subunits in length, 12 to 30 subunits in length, 15 to 30 subunits in length, 18 to 24 subunits in length, 19 to 22 subunits in length, or 20 subunits in length. Antisense oligonucleotides targeted to a PCSK9 nucleic acid may be 8 to 80 nucleotides in length, 12 to 50 nucleotides in length, 12 to 30 nucleotides in length, 15 to 30 nucleotides in length, 18 to 24 nucleotides in length, 19 to 22 nucleotides in length, or 20 nucleotides in length.

Moreover, the antisense compound may be a gapmer antisense oligonucleotide. The gapmer antisense oligonucleotide may comprise a gap segment of ten 2′-deoxynucleotides positioned between wing segments of five 2′-MOE nucleotides. The gapmer antisense oligonucleotide may be a gap-widened antisense oligonucleotide, comprising a gap segment of fourteen 2′-deoxynucleotides positioned between wing segments of three 2′-MOE nucleotides.

Antisense compounds, including antisense oligonucleotides, targeted to a PCSK9 nucleic acid may have at least one modified internucleoside linkage. Additionally, each internucleoside linkage may be a phosphorothioate internucleoside linkage. Each cytosine may be a 5-methyl cytosine.

Antisense compounds, including antisense oligonucleotides, targeted to a PCSK9 nucleic acid may have at least one modified nucleoside. In certain embodiments, a modified nucleoside is a sugar-modified nucleoside. In certain such embodiments, the sugar-modified nucleosides can further comprise a natural or modified heterocyclic base moiety and/or a natural or modified internucleoside linkage and may include further modifications independent from the sugar modification. In certain embodiments, a sugar modified nucleoside is a 2′-modified nucleoside, wherein the sugar ring is modified at the 2′ carbon from natural ribose or 2′-deoxy-ribose.

In certain embodiments, a 2′ modified nucleoside has a 2′-F, 2′-OCH₂ (2′-OMe) or a 2′-O(CH₂)₂—OCH₃ (2′-O-methoxyethyl or 2′-MOE) substituent group

In certain embodiments, a 2′-modified nucleoside has a bicyclic sugar moiety. In certain such embodiments, the bicyclic sugar moiety is a D sugar in the alpha configuration. In certain such embodiments, the bicyclic sugar moiety is a D sugar in the beta configuration. In certain such embodiments, the bicyclic sugar moiety is an L sugar in the alpha configuration. In certain such embodiments, the bicyclic sugar moiety is an L sugar in the beta configuration.

In certain embodiments, the bicyclic sugar moiety comprises a bridge group between the 2′ and the 4′-carbon atoms. In certain such embodiments, the bridge group comprises from 1 to 8 linked biradical groups. In certain embodiments, the bicyclic sugar moiety comprises from 1 to 4 linked biradical groups. In certain embodiments, the bicyclic sugar moiety comprises 2 or 3 linked biradical groups. In certain embodiments, the bicyclic sugar moiety comprises 2 linked biradical groups. In certain embodiments, a linked biradical group is selected from —O—, —S—, —N(R1)-, —C(R1)(R2)-, —C(R1)=C(R1)-, —C(R1)=N—, —C(═NR1)-, —Si(R1)(R2)-, —S(═O)2-, —S(═O)—, —C(═O)— and —C(═S)—; where each R1 and R2 is, independently, H, hydroxyl, C1-C12 alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkynyl, C5-C20 aryl, substituted C5-C20 aryl, a heterocycle radical, a substituted hetero-cycle radical, heteroaryl, substituted heteroaryl, C5-C7 alicyclic radical, substituted C5-C7 alicyclic radical, halogen, substituted oxy (—O—), amino, substituted amino, azido, carboxyl, substituted carboxyl, acyl, substituted acyl, CN, thiol, substituted thiol, sulfonyl (S(═O)2-H), substituted sulfonyl, sulfoxyl (S(═O)—H) or substituted sulfoxyl; and each substituent group is, independently, halogen, C1-C12 alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkynyl, amino, substituted amino, acyl, substituted acyl, C1-C12 aminoalkyl, C1-C12 aminoalkoxy, substituted C1-C12 aminoalkyl, substituted C1-C12 aminoalkoxy or a protecting group.

In some embodiments, the bicyclic sugar moiety is bridged between the 2′ and 4′ carbon atoms with a biradical group selected from —O—(CH2)p-, —O—CH2-, —O—CH2CH2-, —O—CH(alkyl)-, —NH—(CH2)p-, —N(alkyl)-(CH2)p-, —O—CH(alkyl)-, —(CH(alkyl))-(CH2)p-, —NH—O—(CH2)p-, —N(alkyl)-O—(CH2)p-, or —O—N(alkyl)-(CH2)p-, wherein p is 1, 2, 3, 4 or 5 and each alkyl group can be further substituted. In certain embodiments, p is 1, 2 or 3.

In one aspect, each of said bridges is, independently, —[C(R1)(R2)]n-, —[C(R1)(R2)]n-O—, —C(R1R2)-N(R1)-O— or —C(R1R2)-O—N(R1)-. In another aspect, each of said bridges is, independently, 4′-(CH2)3-2′, 4′-(CH2)2-2′, 4′-CH2-O-2′, 4′-(CH2)2-O-2′, 4′-CH2-O—N(R1)-2′ and 4′-CH2-N(R1)-O-2′- wherein each R1 is, independently, H, a protecting group or C1-C12 alkyl.

The present disclosure further discloses subsets of the antisense compounds above that show superior properties relating to pharmacodynamics and/or pharmacokinetics, among other properties. Exemplary antisense compounds reduce PCSK9 expression in a cultured cell model system, e.g., Hep3B cells, cynomolgus monkey hepatocytes, or human primary hepatocytes. In particular embodiments, antisense compounds reduce PCSK9 expression in a dose-dependent manner in a cell culture system, where the antisense compounds have an IC₅₀ in the nanomolar range.

Exemplary antisense compounds also reduce the expression of a PCSK9 nucleic acid in an animal model, such as a transgenic mouse that expresses a human PCSK9 nucleic acid (HuPCSK9Tg mice). In particular embodiments, antisense compounds include those that reduce PCSK9 express in an animal model with a physiology that correlates closely with humans, such as monkey, e.g., the cynomolgus monkey.

Exemplary antisense compounds display minimal side effects. Side effects include responses to the administration of the antisense compound unrelated to the targeting of a PCSK9 nucleic acid, such as an inflammatory response in the host individual. Exemplary antisense compounds are well tolerated by the host individual. Tolerability may be determined though histological analysis of various cell types, and includes examination of such markers as cytoplasmic swelling from multifocal apoptosis in liver cells, follicular hyperplasia in spleen cells, macrophage infiltration, and the like. In particular embodiments, antisense compounds produce minimal signs of host intolerance.

Exemplary antisense compounds further display favorable pharmacokinetics. In particular embodiments, antisense compounds accumulate at a relatively high ratio in the liver versus other sensitive organs, such as kidneys. In particular embodiments, antisense compounds exhibit relatively high half-lives in relevant biological fluids or tissues, e.g., liver tissue, reflecting higher stability and resistance to nucleases, for example.

In particular embodiments, antisense compounds target identical sequences in human and animal PCSK9 sequences. In other embodiments, antisense compounds target PCSK9 sequences without a known single nucleotide polymorphism (SNP), have a high relative G-content, have minimal secondary structure, have greatly reduced effects on the host's serum chemistry, body weight, or histopathology, compared to other antisense compounds, and/or induce a minimal adverse immunohistocompatibility reaction.

In a particular embodiment, the antisense compound is selected from the group of Isis 405881, Isis 399819, Isis 395165, Isis 405879, Isis 406008, Isis 405891, Isis 395186, Isis 405988, Isis 405994, Isis 406023, Isis 395187, Isis 395185, Isis 406033, Isis 405923, Isis 399900, Isis 405995, Isis 405991, Isis 406005, Isis 399793, and Isis 395152. The aforementioned antisense compounds effectively repress PCSK9 expression in a Hep3B cell culture system. In another embodiment, antisense compounds effectively inhibit PCSK9 expression with a low IC₅₀ in a battery of cell culture systems. Exemplary antisense compounds include those selected from the group of Isis 395165, Isis 395185, Isis 395186, Isis 395187, Isis 405879, Isis 405881, Isis 405891, Isis 405988, Isis 405994, and Isis 406008. In yet another embodiment, antisense compounds exhibit minimal or no adverse histopathological effects when administered at particularly high doses to an individual. Exemplary antisense compounds include those selected from the group consisting of Isis 395185, Isis 395186, Isis 395187, Isis 405879, Isis 405891, and Isis 405988. In yet another embodiment, antisense compounds affect therapeutic end-points, e.g., reduction of plasma LDL-C, liver TG, or liver PCSK9 mRNA, in an animal model. Exemplary antisense compounds include Isis 405879. In yet another embodiment, antisense compounds display combinations of the characteristics above and reduce liver PCSK9 mRNA expression in an animal model with high efficiency, e.g., Isis 405879.

DEFINITIONS

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by on of skill in the art to which the invention(s) belong. Unless specific definitions are provided, the nomenclature utilized in connection with, and the procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well known and commonly used in the art. Standard techniques may be used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and delivery, and treatment of subjects. Certain such techniques and procedures may be found for example in “Antisense Drug Technology: Principles, Strategies, and Applications.” by Stanley Crooke, Boca Raton: Taylor & Francis Group, 2008; “Carbohydrate Modifications in Antisense Research” Edited by Sangvi and Cook, American Chemical Society, Washington D.C., 1994; and “Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton, Pa., 18th edition, 1990; and which is hereby incorporated by reference for any purpose. Where permitted, all patents, patent applications, published applications and publications, GENBANK sequences, websites and other published materials referred to throughout the entire disclosure herein, unless noted otherwise, are incorporated by reference in their entirety. All of the GENBANK® Accession Nos. along with their associated sequence and structural data pertaining to such sequences including gene organization and structural elements and SNP information that may be found in sequence databases such as the National Center for Biotechnology Information (NCBI) are incorporated herein by reference in their entirety. In the event that there is a plurality of definitions for terms herein, those in this section prevail. Where reference is made to a URL or other such identifier or address, it is understood that such identifiers can change and particular information on the internet can come and go, but equivalent information can be found by searching the internet. Reference thereto evidences the availability and public dissemination of such information.

A “pharmaceutical composition” means a mixture of substances suitable for administering to an individual. For example, a pharmaceutical composition may comprise an antisense oligonucleotide and a sterile aqueous solution.

“Administering” means providing a pharmaceutical agent to an individual, and includes, but is not limited to administering by a medical professional and self-administering.

“Individual” means a human or non-human animal selected for treatment or therapy.

“Duration” means the period of time during which an activity or event continues. In certain embodiments, the duration of treatment is the period of time during which doses of a pharmaceutical agent are administered.

“Parenteral administration” means administration through injection or infusion. Parenteral administration includes, but is not limited to, subcutaneous administration, intravenous administration, or intramuscular administration.

“Subcutaneous administration” means administration just below the skin. “Intravenous administration” means administration into a vein.

“Dose” means a specified quantity of a pharmaceutical agent provided in a single administration. In certain embodiments, a dose may be administered in two or more boluses, tablets, or injections. For example, in certain embodiments, where subcutaneous administration is desired, the desired dose requires a volume not easily accommodated by a single injection. In such embodiments, two or more injections may be used to achieve the desired dose. In certain embodiments, a dose may be administered in two or more injections to minimize injection site reaction in a individual.

“Dosage unit” means a form in which a pharmaceutical agent is provided. In certain embodiments, a dosage unit is a vial containing lyophilized antisense oligonucleotide. In certain embodiments, a dosage unit is a vial containing reconstituted antisense oligonucleotide.

“Pharmaceutical agent” means a substance provides a therapeutic benefit when administered to a individual. For example, in certain embodiments, an antisense oligonucleotide targeted to PCSK9 is pharmaceutical agent.

“Diluent” means an ingredient in a composition that lacks pharmacological activity, but is pharmaceutically necessary or desirable. For example, in drugs that are injected the diluent may be a liquid, e.g., saline solution.

“Active pharmaceutical ingredient” means the substance in a pharmaceutical composition that provides a desired effect.

“Pharmaceutically acceptable carrier” means a medium or diluent that does not interfere with the structure of the oligonucleotide. Certain of such carriers enable pharmaceutical compositions to be formulated as, for example, tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspension and lozenges for the oral ingestion by a subject.

“Therapeutically effective amount” means an amount of a pharmaceutical agent that provides a therapeutic benefit to an individual. In certain embodiments, a therapeutically effective amount of antisense compound targeted to a PCSK9 nucleic acid is an amount that decreases LDL-C in the individual.

“Hypercholesterolemia” means a condition characterized by elevated serum cholesterol.

“Hyperlipidemia” means a condition characterized by elevated serum lipids.

“Hypertriglyceridemia” means a condition characterized by elevated serum triglyceride levels.

“Non-familial hypercholesterolemia” means a condition characterized by elevated serum cholesterol that is not the result of a single gene mutation.

“Polygenic hypercholesterolemia” means a condition characterized by elevated cholesterol that results from the influence of a variety of genetic factors. In certain embodiments, polygenic hypercholesterolemia may be exacerbated by dietary intake of lipids.

“Familial hypercholesterolemia (FH)” means an autosomal dominant metabolic disorder characterized by a mutation in the LDL-receptor (LDL-R) gene, markedly elevated LDL-C and premature onset of atherosclerosis. A diagnosis of familial hypercholesterolemia is made when an individual meets one or more of the following criteria: genetic testing confirming 2 mutated LDL-receptor genes; genetic testing confirming one mutated LDL-receptor gene; document history of untreated serum LDL-cholesterol greater than 500 mg/dL; tendinous and/or cutaneous xanthoma prior to age 10 years; or, both parents have documented elevated serum LDL-cholesterol prior to lipid-lowering therapy consistent with heterozygous familial hypercholesterolemia.

“Homozygous familial hypercholesterolemia” or “HoFH” means a condition characterized by a mutation in both maternal and paternal LDL-R genes.

“Heterozygous familial hypercholesterolemia” or “HeFH” means a condition characterized by a mutation in either the maternal or paternal LDL-R gene.

“Mixed dyslipidemia” means a condition characterized by elevated serum cholesterol and elevated serum triglycerides.

“Diabetic dyslipidemia” or “Type II diabetes with dyslipidemia” means a condition characterized by Type II diabetes, reduced HDL-C, elevated serum triglycerides, and elevated small, dense LDL particles.

“CHD risk equivalents,” means indicators of clinical atherosclerotic disease that confer a high risk for coronary heart disease, and include clinical coronary heart disease, symptomatic carotid artery disease, peripheral arterial disease, and/or abdominal aortic aneurysm.

“Metabolic syndrome” means a condition characterized by a clustering of lipid and non-lipid cardiovascular risk factors of metabolic origin. In certain embodiments, metabolic syndrome is identified by the presence of any 3 of the following factors: waist circumference of greater than 102 cm in men or greater than 88 cm in women; serum triglyceride of at least 150 mg/dL; HDL-C less than 40 mg/dL in men or less than 50 mg/dL in women; blood pressure of at least 130/85 mmHg; and fasting glucose of at least 110 mg/dL.

“Non-alcoholic fatty liver disease (NAFLD)” means a condition characterized by fatty inflammation of the liver that is not due to excessive alcohol use (for example, alcohol consumption of over 20 g/day). In certain embodiments, NAFLD is related to insulin resistance and the metabolic syndrome.

“Non-alcoholic steatohepatitis (NASH)” means a condition characterized by inflammation and the accumulation of fat and fibrous tissue in the liver, that is not due to excessive alcohol use. NASH is an extreme form of NAFLD.

“Major risk factors” mean factors that contribute to a high risk for coronary heart disease, and include without limitation cigarette smoking, hypertension, low HDL-C, family history of coronary heart disease, and age.

“CHD risk factors” mean CHD risk equivalents and major risk factors.

“Coronary heart disease (CHD)” means a narrowing of the small blood vessels that supply blood and oxygen to the heart, which is often a result of atherosclerosis.

“Reduced coronary heart disease risk” means a reduction in the likelihood that an individual will develop coronary heart disease. In certain embodiments, a reduction in coronary heart disease risk is measured by an improvement in one or more CHD risk factors, for example, a decrease in LDL-C levels.

“Atherosclerosis” means a hardening of the arteries affecting large and medium-sized arteries and is characterized by the presence of fatty deposits. The fatty deposits are called “atheromas” or “plaques,” which consist mainly of cholesterol and other fats, calcium and scar tissue, and damage the lining of arteries.

“History of coronary heart disease” means the occurrence of clinically evident coronary heart disease in the medical history of an individual or a individual's family member.

“Early onset coronary heart disease” means a diagnosis of coronary heart disease prior to age 50.

“Statin intolerant individual” means an individual who as a result of statin therapy experiences one or more of creatine kinase increases, liver function test abnormalities, muscle aches, or central nervous system side effects.

“Efficacy” means the ability to produce a desired effect. For example, efficacy of a lipid-lowering therapy may be reduction in the concentration of one or more of LDL-C, VLDL-C, IDL-C, non-HDL-C, ApoB, lipoprotein(a), or triglycerides.

“Acceptable safety profile” means a pattern of side effects that is within clinically acceptable limits.

“Side effects” means physiological responses attributable to a treatment other than desired effects. In certain embodiments, side effects include, without limitation, injection site reactions, liver function test abnormalities, renal function abnormalities, liver toxicity, renal toxicity, central nervous system abnormalities, and myopathies. For example, increased aminotransferase levels in serum may indicate liver toxicity or liver function abnormality. For example, increased bilirubin may indicate liver toxicity or liver function abnormality.

“Injection site reaction” means inflammation or abnormal redness of skin at a site of injection in an individual.

“Individual compliance” means adherence to a recommended or prescribed therapy by an individual.

“Lipid-lowering therapy” means a therapeutic regimen provided to an individual to reduce one or more lipids in a individual. In certain embodiments, a lipid-lowering therapy is provide to reduce one or more of ApoB, total cholesterol, LDL-C, VLDL-C, IDL-C, non-HDL-C, triglycerides, small dense LDL particles, and Lp(a) in a individual.

“Lipid-lowering agent” means a pharmaceutical agent provided to an individual to achieve a lowering of lipids in the individual. For example, in certain embodiments, a lipid-lowering agent is provided to an individual to reduce one or more of ApoB, LDL-C, total cholesterol, and triglycerides.

“LDL-C target” means an LDL-C level that is desired following lipid-lowering therapy.

“Comply” means the adherence with a recommended therapy by a individual.

“Recommended therapy” means a therapeutic regimen recommended by a medical professional for the treatment, amelioration, or prevention of a disease.

“Low LDL-receptor activity” means LDL-receptor activity that is not sufficiently high to maintain clinically acceptable levels of LDL-C in the bloodstream.

“Cardiovascular outcome” means the occurrence of major adverse cardiovascular events.

“Improved cardiovascular outcome” means a reduction in the occurrence of major adverse cardiovascular events, or the risk thereof. Examples of major adverse cardiovascular events include, without limitation, death, reinfarction, stroke, cardiogenic shock, pulmonary edema, cardiac arrest, and atrial dysrhythmia.

“Surrogate markers of cardiovascular outcome” means indirect indicators of cardiovascular events, or the risk thereof. For example, surrogate markers of cardiovascular outcome include carotid intimal media thickness (CIMT). Another example of a surrogate marker of cardiovascular outcome includes atheroma size. Atheroma size may be determined by intravascular ultrasound (IVUS). Surrogate markers also include increased HDL-cholesterol, or any combination of the markers above.

“Increased HDL-C” means an increase in serum HDL-C in an individual over time.

“Lipid-lowering” means a reduction in one or more serum lipids in an individual over time.

“Co-administration” means administration of two or more pharmaceutical agents to a individual. The two or more pharmaceutical agents may be in a single pharmaceutical composition, or may be in separate pharmaceutical compositions. Each of the two or more pharmaceutical agents may be administered through the same or different routes of administration. Co-administration encompasses administration in parallel or sequentially.

“Administered concomitantly” refers to the administration of two agents at the same therapeutic time frame, in any manner in which the pharmacological effects of both are manifest in the patient at the same time. Concomitant administration does not require that both agents be administered in a single pharmaceutical composition, in the same dosage form, or by the same route of administration.

“Therapeutic lifestyle change” means dietary and lifestyle changes intended to lower cholesterol and reduce the risk of developing heart disease, and includes recommendations for dietary intake of total daily calories, total fat, saturated fat, polyunsaturated fat, monounsaturated fat, carbohydrate, protein, cholesterol, insoluble fiber, as well as recommendations for physical activity.

“Statin” means a pharmaceutical agent that inhibits the activity of HMG-CoA reductase.

“HMG-CoA reductase inhibitor” means a pharmaceutical agent that acts through the inhibition of the enzyme HMG-CoA reductase.

“Cholesterol absorption inhibitor” means a pharmaceutical agent that inhibits the absorption of exogenous cholesterol obtained from diet.

“LDL apheresis” means a form of apheresis by which LDL-C is removed from blood. Typically, a individual's blood is removed from a vein, and separated into red cells and plasma. LDL-C is filtered out of the plasma prior to return of the plasma and red blood cells to the individual.

“MTP inhibitor” means a pharmaceutical agent that inhibits the enzyme microsomal triglyceride transfer protein.

“Low density lipoprotein-cholesterol (LDL-C)” means cholesterol carried in low density lipoprotein particles. Concentration of LDL-C in serum (or plasma) is typically quantified in mg/dL or nmol/L. “Serum LDL-C” and “plasma LDL-C” mean LDL-C in the serum and plasma, respectively.

“Very low density lipoprotein-cholesterol (VLDL-C)” means cholesterol associated with very low density lipoprotein particles. Concentration of VLDL-C in serum (or plasma) is typically quantified in mg/dL or nmol/L. “Serum VLDL-C” and “plasma VLDL-C” mean VLDL-C in the serum or plasma, respectively.

“Intermediate low density lipoprotein-cholesterol (IDL-C)” means cholesterol associated with intermediate density lipoprotein. Concentration of IDL-C in serum (or plasma) is typically quantified in mg/dL or nmol/L. “Serum IDL-C” and “plasma IDL-C” mean IDL-C in the serum or plasma, respectively.

“Non-high density lipoprotein-cholesterol (Non-HDL-C)” means cholesterol associated with lipoproteins other than high density lipoproteins, and includes, without limitation, LDL-C, VLDL-C, and IDL-C.

“High density lipoprotein-C (HDL-C)” means cholesterol associated with high density lipoprotein particles. Concentration of HDL-C in serum (or plasma) is typically quantified in mg/dL or nmol/L. “Serum HDL-C” and “plasma HDL-C” mean HDL-C in the serum and plasma, respectively.

“Total cholesterol” means all types of cholesterol, including, but not limited to, LDL-C, HDL-C, IDL-C and VLDL-C. Concentration of total cholesterol in serum (or plasma) is typically quantified in mg/dL or nmol/L.

“Lipoprotein(a)” or “Lp(a)” means a lipoprotein particle that is comprised of LDL-C, an apolipoprotein(a) particle, and an apolipoproteinB-100 particle.

“ApoA1” means apolipoprotein-A1 protein in serum. Concentration of ApoA1 in serum is typically quantified in mg/dL or nmol/L.

“ApoB:ApoA1 ratio” means the ratio of ApoB concentration to ApoA1 concentration.

“ApoB-containing lipoprotein” means any lipoprotein that has apolipoprotein B as its protein component, and is understood to include LDL, VLDL, IDL, and lipoprotein(a).

“Small LDL particle” means a subclass of LDL particles characterized by a smaller, denser size compared to other LDL particles. In certain embodiments, large LDL particles are 23-27 nm in diameter. In certain embodiments, intermediate LDL particles are 21.2-23 nm in diameter. In certain embodiments, small LDL particles are 18-21.2 nm in diameter. In certain embodiments, particle size is measured by nuclear magnetic resonance analysis.

“Small VLDL particle” means a subclass of VLDL particles characterized by a smaller, denser size compared to other VLDL particles. In certain embodiments, large VLDL particles are greater than 60 nm in diameter. In certain embodiments, medium VLDL particles are 35-60 nm in diameter. In certain embodiments, small VLDL particles are 27-35 nm in diameter. In certain embodiments, particle size is measured by nuclear magnetic resonance analysis.

“Triglycerides” means lipids that are the triesters of glycerol. “Serum triglycerides” mean triglycerides present in serum. “Liver triglycerides” mean triglycerides present in liver tissue.

“Serum lipids” mean cholesterol and triglycerides in the serum.

“Elevated total cholesterol” means total cholesterol at a concentration in an individual at which lipid-lowering therapy is recommended, and includes, without limitation, elevated LDL-C”, “elevated VLDL-C,” “elevated IDL-C,” and “elevated non-HDL-C.” In certain embodiments, total cholesterol concentrations of less than 200 mg/dL, 200-239 mg/dL, and greater than 240 mg/dL are considered desirable, borderline high, and high, respectively. In certain embodiments, LDL-C concentrations of 100 mg/dL, 100-129 mg/dL, 130-159 mg/dL, 160-189 mg/dL, and greater than 190 mg/dL are considered optimal, near optimal/above optimal, borderline high, high, and very high, respectively.

“Elevated triglyceride” means concentrations of triglyceride in the serum or liver at which lipid-lowering therapy is recommended, and includes “elevated serum triglyceride” and “elevated liver triglyceride.” In certain embodiments, serum triglyceride concentration of 150-199 mg/dL, 200-499 mg/dL, and greater than or equal to 500 mg/dL is considered borderline high, high, and very high, respectively.

“Elevated small LDL particles” means a concentration of small LDL particles in an individual at which lipid-lowering therapy is recommended.

“Elevated small VLDL particles” means a concentration of small VLDL particles in an individual at which lipid-lowering therapy is recommended.

“Elevated lipoprotein(a)” means a concentration of lipoprotein(a) in an individual at which lipid-lowering therapy is recommended.

“Low HDL-C” means a concentration of HDL-C in an individual at which lipid-lowering therapy is recommended. In certain embodiments lipid-lowering therapy is recommended when low HDL-C is accompanied by elevations in non-HDL-C and/or elevations in triglyceride. In certain embodiments, HDL-C concentrations of less than 40 mg/dL are considered low. In certain embodiments, HDL-C concentrations of less than 50 mg/dL are considered low.

“ApoB” means apolipoprotein B-100 protein. Concentration of ApoB in serum (or plasma) is typically quantified in mg/dL or nmol/L. “Serum ApoB” and “plasma ApoB” mean ApoB in the serum and plasma, respectively.

“LDL/HDL ratio” means the ratio of LDL-C to HDL-C.

“Oxidized-LDL” or “Ox-LDL-C” means LDL-C that is oxidized following exposure to free radicals.

“Individual having elevated LDL-C levels” means an individual who has been identified by a medical professional (e.g., a physician) as having LDL-C levels near or above the level at which therapeutic intervention is recommended, according to guidelines recognized by medical professionals. Such an individual may also be considered “in need of treatment” to decrease LDL-C levels.

“Individual having elevated apoB-100 levels” means an individual who has been identified as having apoB-100 levels near or above the level at which therapeutic intervention is recommended, according to guidelines recognized by medical professionals. Such an individual may also be considered “in need of treatment” to decrease apoB-100 levels.

“Treatment of elevated LDL-C levels” means administration of an antisense compound targeted to a PCSK9 nucleic acid to an individual having elevated LDL-C levels.

“Treatment of atherosclerosis” means administration of an antisense compound targeted to a PCSK9 nucleic acid to an individual who, based upon a physician's assessment, has or is likely to have atherosclerosis. “Prevention of atherosclerosis” means administration of an antisense compound targeted to a PCSK9 nucleic acid to an individual who, based upon a physician's assessment, is susceptible to atherosclerosis.

“Ameliorate” means to make better or improve a detrimental condition in an individual.

“Target nucleic acid,” “target RNA,” “target RNA transcript” and “nucleic acid target” all mean any nucleic acid capable of being targeted by antisense compounds.

“PCSK9 nucleic acid” means any nucleic acid encoding PCSK9. For example, in certain embodiments, a PCSK9 nucleic acid includes, without limitation, a DNA sequence encoding PCSK9, an RNA sequence transcribed from DNA encoding PCSK9, and an mRNA sequence encoding PCSK9. “PCSK9 mRNA” means an mRNA encoding a PCSK9 protein.

“Targeting” means the process of design and selection of an antisense compound that will specifically hybridize to a target nucleic acid and induce a desired effect.

“Targeted” means having a nucleobase sequence that will allow specific hybridization of an antisense compound to a target nucleic acid to induce a desired effect. In certain embodiments, a desired effect is reduction of a target nucleic acid. In certain such embodiments, a desired effect is reduction of PCSK9 mRNA.

“Antisense inhibition” means reduction of a target nucleic acid levels in the presence of an antisense compound complementary to a target nucleic acid compared to target nucleic acid levels in the absence of the antisense compound.

“Target region” means a fragment of a target nucleic acid to which one or more antisense compounds is targeted.

“Target segment” means the sequence of nucleotides of a target nucleic acid to which an antisense compound is targeted. “5′ target site” refers to the 5′-most nucleotide of a target segment. “3′ target site” refers to the 3′-most nucleotide of a target segment.

“Active target region” means a target region to which one or more active antisense compounds is targeted. “Active antisense compounds” means antisense compounds that reduce target nucleic acid levels.

“Hybridization” means the annealing of complementary nucleic acid molecules. In certain embodiments, complementary nucleic acid molecules include, but are not limited to, an antisense compound and a nucleic acid target. In certain such embodiments, complementary nucleic acid molecules include, but are not limited to, an antisense oligonucleotide and a nucleic acid target

“Stringent hybridization conditions” means conditions under which a nucleic acid molecule, such as an antisense compound, will hybridize to a target nucleic acid sequence, but to a minimal number of other sequences.

“Specifically hybridizable” means an antisense compound that hybridizes to a target nucleic acid to induce a desired effect, while exhibiting minimal or no effects on non-target nucleic acids.

“Complementarity” means the capacity for pairing between nucleobases of a first nucleic acid and a second nucleic acid.

“Fully complementary” means each nucleobase of a first nucleic acid is capable of pairing with each nucleobase of a second nucleic acid. In certain embodiments, a first nucleic acid is an antisense compound and a target nucleic acid is a second nucleic acid. In certain such embodiments, an antisense oligonucleotide is a first nucleic acid and a target nucleic acid is a second nucleic acid.

“Non-complementary nucleobase” means a nucleobase of first nucleic acid that is not capable of pairing with the corresponding nucleobase of a target nucleic acid. “Mismatch” a nucleobase of first nucleic acid that is not capable of pairing with the corresponding nucleobase of a target nucleic acid.

“Portion” means a defined number of contiguous (i.e., linked) nucleobases. In certain embodiments, a portion is a defined number of contiguous nucleobases of a target nucleic acid. In certain embodiments, a portion is a defined number of contiguous nucleobases of an antisense compound.

“Oligomeric compound” means a polymer of linked monomeric subunits which is capable of hybridizing to at least a region of an RNA molecule.

“Antisense compound” means an oligomeric compound that is capable of undergoing hybridization to a target nucleic acid through hydrogen bonding.

“Oligonucleotide” means a polymer of linked nucleosides each of which can be modified or unmodified, independent one from another.

“Oligonucleoside” means an oligonucleotide in which the internucleoside linkages do not contain a phosphorus atom.

“Unmodified nucleotide” means a nucleotide composed of naturally occurring nucleobases, sugar moieties and internucleoside linkages. In certain embodiments, an unmodified nucleotide is an RNA nucleotide (i.e., β-D-ribonucleosides) or a DNA nucleotide (i.e., β-D-deoxyribonucleoside).

“Antisense oligonucleotide” means a single-stranded oligonucleotide having a nucleobase sequence that permits hybridization to a corresponding region of a target nucleic acid.

“Motif” means the pattern of unmodified and modified nucleosides in an antisense compound.

“Chimeric antisense compound” means an antisense compound that has at least 2 chemically distinct regions, each region having a plurality of subunits.

A “gapmer” means an antisense compound in which an internal position having a plurality of nucleotides that supports RNaseH cleavage is positioned between external regions having one or more nucleotides that are chemically distinct from the nucleosides of the internal region.

A “gap segment” means the plurality of nucleotides that make up the internal region of a gapmer.

A “wing segment” means the external region of a gapmer.

“Gap-widened” means an antisense compound has a gap segment of 12 or more contiguous 2′-deoxyribonucleotides positioned between 5′ and 3′ wing segments having from one to six nucleotides having modified sugar moieties.

“Nucleoside” means a nucleobase linked to a sugar.

“Nucleobase” means a heterocyclic base moiety capable of pairing with a base of another nucleic acid.

“Nucleotide” means a nucleoside having a phosphate group covalently linked to the sugar portion of the nucleoside.

“Nucleobase sequence” means the order of contiguous nucleobases independent of any sugar, linkage, and/or nucleobase modification.

“Modified nucleotide” means a nucleotide having, independently, a modified sugar moiety, modified internucleoside linkage, or modified nucleobase.

“Modified nucleoside” means a nucleotide having, independently, a modified sugar moiety or modified nucleobase.

“Contiguous nucleobases” means nucleobases immediately adjacent to each other.

“Modified oligonucleotide” means an oligonucleotide comprising a modified internucleoside linkage, a modified sugar, and/or a modified nucleobase.

“Single-stranded modified oligonucleotide” means a modified oligonucleotide which is not hybridized to a complementary strand.

“Internucleoside linkage” refers to the chemical bond between nucleosides.

“Linked nucleoside” means adjacent nucleosides which are bonded together.

“Linked deoxynucleoside” means a nucleic acid base (A, G, C, T, U) substituted by deoxyribose linked by a phosphate ester to form a nucleotide.

“Naturally occurring internucleoside linkage” means a 3′ to 5′ phosphodiester linkage.

“Modified internucleoside linkage” means substitution and/or any change from a naturally occurring internucleoside bond. In certain instances, the modified internucleoside linkage refers to a phosphodiester internucleoside bond.

“Phosphorothioate linkage” means a linkage between nucleosides where the phosphodiester bond is modified by replacing one of the non-bridging oxygen atoms with a sulfur atom. A phosphorothioate linkage is a modified internucleoside linkage.

“Modified sugar moiety” means substitution and/or any change from a natural sugar moiety. For the purposes of this disclosure, a “natural sugar moiety” is a sugar moiety found in DNA (2′-H) or RNA (2′-OH).

“Modified sugar” refers to a substitution and/or any change from a natural sugar.

“Bicyclic sugar” means a furosyl ring modified by the bridging of the two non-geminal ring atoms. A bicyclic sugar is a modified sugar.

“Modified nucleobase” means any nucleobase other than adenine, cytosine, guanine, thymidine, or uracil. An “unmodified nucleobase” means the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) and uracil (U).

“Modified sugar moiety” means a sugar moiety having a substitution and/or any change from a natural sugar moiety.

“Natural sugar moiety” means a sugar moiety found in DNA (2′-H) or RNA (2′-OH).

“2′-O-methoxyethyl sugar moiety” means a 2′-substituted furosyl ring having a 2′-O(CH2)₂-OCH₃ (2′-O-methoxyethyl or 2′-MOE) substituent group.

“2′-O-methoxyethyl nucleotide” means a nucleotide comprising a 2′-O-methoxyethyl modified sugar moiety.

“2′-O-methoxyethyl” refers to an O-methoxy-ethyl modification of the 2′ position of a furosyl ring. A 2′-O-methoxyethyl modified sugar is a modified sugar.

“Bicyclic nucleic acid sugar moiety” means a furosyl ring modified by the bridging of two non-geminal ring atoms.

“5-methylcytosine” means a cytosine modified with a methyl group attached to the 5′ position. A 5-methylcytosine is a modified nucleobase.

“Prodrug” means a therapeutic agent that is prepared in an inactive form that is converted to an active form (i.e., drug) within the body or cells thereof by the action of endogenous enzymes or other chemicals and/or conditions.

“Pharmaceutically acceptable salts” means physiologically and pharmaceutically acceptable salts of antisense compounds, i.e., salts that retain the desired biological activity of the parent oligonucleotide and do not impart undesired toxicological effects thereto.

“Salts” refers to physiologically and pharmaceutically acceptable salts of antisense compounds, i.e., salts that retain the desired biological activity of the parent oligonucleotide and do not impart undesired toxicological effects thereto.

“Cures” means a method or course that restores health or a prescribed treatment for an illness.

“Slows progression” means decrease in the development of the said disease.

“Cap structure” or “terminal cap moiety” means chemical modifications, which have been incorporated at either terminus of an antisense compound.

“ISIS 301012” means a lipid-lowering agent that is an antisense oligonucleotide having the sequence “GCCTCAGTCTGCTTCGCACC” (SEQ ID NO: 457), where each internucleoside linkage is a phosphorothioate internucleoside linkage, each cytosine is a 5-methylcytosine, nucleotides 6-15 are 2′-deoxynucleotides, and nucleotides 1-5 and 16-20 are 2′-O-methoxyethyl nucleotides. ISIS 301012 does not target PCSK9. It is used herein as a non-PCSK9 control.

BRIEF DESCRIPTION OF THE FIGURES

The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

FIG. 1 depicts the location of PCSK9 mRNA target sequences of exemplary antisense compounds.

FIG. 2 depicts the results of a five-point dose response experiment with exemplary antisense compounds in HEK-293 cells. The antisense compounds are identified by Isis number. The bars from left to right represent PCSK9 expression in relative units at 0 nM, 25 nM, 50 nM, 100 nM, 200 nM, and 300 nM antisense oligonucleotides.

FIG. 3 depicts the results of a five-point dose response experiment with exemplary antisense compounds in cyno primary hepatocytes. The antisense compounds are identified by Isis number. The bars from left to right represent PCSK9 expression in relative units at 300 nM, 150 nM, 50 nM, 25 nM, and 10 nM antisense oligonucleotides. Control values are shown in the rightmost two bars.

FIG. 4 depicts a reduction of murine PCSK9 mRNA expression (relative units) in the liver of h-apoB/CETP transgenic mice when administered Isis 394816. Isis 394816 was administered by intraperitoneal injection for 6 weeks at a dosing regimen of 20 mg/kg/wk or 50 mg/kg/wk. Isis 329993 was administered at 50 mg/kg/wk as a control.

FIG. 5, Panel A, depicts inhibition of plasma LDL-C (mg/dL) in mice administered Isis 394816 at 20 mg/kg/wk or 50 mg/kg/wk. FIG. 5, Panel B, depicts inhibition of liver TG (mg/gram) in mice administered Isis 394816 at 20 mg/kg/wk or 50 mg/kg/wk.

FIG. 6 depicts the dose-dependent inhibition (relative units) of liver PCSK9 mRNA, plasma LDL-C, and liver TG in response to the indicated doses of Isis 394816.

FIG. 7 depicts a correlation between LDL-C lowering and PCSK9 mRNA inhibition, using the data from FIG. 6.

FIG. 8 depicts the accumulation of exemplary antisense compounds in rat tissues (μg/g tissue) following administration of antisense compounds. The antisense compounds are identified by Isis number. The bars from left to right represent antisense compound accumulation in kidney and liver.

FIG. 9 depicts the accumulation of the identified antisense compounds in tissues (μg/g tissue) following administration to a cyno monkey. Accumulation at each dosing concentration (15 mg/kg and 30 mg/kg) is shown in cyno kidney (left side bar) and cyno liver (right side bar).

FIG. 10 depicts PCSK9 mRNA levels (relative units) following administration of the identified antisense compounds to a cyno monkey. Results are shown at each dosing concentration (15 mg/kg and 30 mg/kg).

FIG. 11 depicts hepatic PCSK9 mRNA levels (relative units) detected by Northern analysis. Cyno monkeys were administered Isis 405879 (“BMS-844421”) at a dose of 15 mg/kg or 30 mg/kg.

FIG. 12 depicts plasma LDL-C levels (mg/dL) following administration of Isis 405879 (labeled “BMS-844421”) at a dose of 15 mg/kg or 30 mg/kg. Left side bars represent baseline levels of LDL-C in control animals; right side bars represent LDL-C following administration of antisense compounds.

FIG. 13 depicts the ratio of plasma HDL/LDL following administration of Isis 405879 (labeled “BMS-844421”) at a dose of 15 mg/kg or 30 mg/kg. Left side bars represent baseline HDL/LDL in control animals; right side bars represent HDL/LDL following administration of antisense compounds.

FIG. 14 depicts the effect of BMS-844421 and control ASOs on liver human PCSK9 mRNA in transgenic mice (6 wk).

FIG. 15 depicts the effect of BMS-844421 and control ASOs on plasma levels of human PCSK9 protein in transgenic mice (6 wk).

FIG. 16 depicts plasma hPCSK9 at baseline (FIG. 16A), 3 wk (FIG. 16B), and 6 wk (FIG. 16C) treatment.

FIG. 17 depicts the BMS-844421 dose response for plasma hPCSK9 levels in transgenic mice at 6 weeks (wk).

FIG. 18 depicts the BMS-844421 dose response for liver mRNA and plasma protein (hPCSK9).

FIG. 19 depicts the dose response for plasma hPCSK9 in male and female transgenic mice.

FIG. 20 depicts the dose response for liver hPCSK9 mRNA in male versus female transgenic mice.

FIG. 21 depicts the effect of the ASOs on liver endogenous PCSK9 mRNA in the transgenic mouse after 6 weeks of administration. The numbers following the hyphens provide the mg ASO/kg body weight dose of each ASO. For example, “844421-30” is the data for BMS-84421 administered to test animals at 30 mg/kg.

FIG. 22 depicts the effect of ASOs on liver endogenous HMGCoA reductase mRNA in the transgenic mouse after 6 weeks of administration.

FIG. 23 depicts a Western blot, using antibodies directed to the transferrin receptor (TR) in red and antibodies directed to LDLR protein in green. LDLR protein concentration is assayed following a 6 wk ASO treatment.

FIG. 24 depicts the quantification of the Western blot shown in FIG. 23. The LDLR band is normalized to the transferrin receptor (TR) band, as observed using anti-LDLR and anti-TR antibodies, respectively.

FIG. 25 depicts the Liver PCSK9 mRNA concentration in cyno monkeys after 13 weeks of treatment. The numbers in parentheses are administered doses in mg/kg.

FIG. 26 depicts the concentration of BMS-844421 in the liver of cyno monkeys after 13 weeks of treatment.

FIG. 27 depicts the average serum LDL-C levels in the cyno monkey study. The numbers in parentheses are administered doses in mg/kg.

FIG. 28A depicts the pharmacodynamic responses in serum lipid assays for the cyno 2.5 and 5 wk ASO studies, as displayed in a scatterplot of individual animal data. FIG. 28B depicts the average ratio of HDL/LDL in BMS-844421-treated cynos at 2.5 and 5 wk. The numbers in parentheses in FIG. 28B are administered doses in mg/kg.

FIG. 29 depicts downregulation of PCSK9 target in cyno monkeys at 13 wk in the toxicology study of liver mRNA and plasma PCSK9.

DETAILED DESCRIPTION

Overview

Elevated levels of LDL-cholesterol (HDL-C) are recognized as a major independent risk factor for coronary heart disease (CHD). Even in individuals undergoing aggressive treatment with currently available cholesterol-lowering agents to reduce LDL-cholesterol (LDL-C) levels, coronary events still occur, and elevated LDL-C levels remain a major risk factor for coronary heart disease in these individuals. Furthermore, many individuals undergoing LDL-lowering therapy do not reach their target LDL-C levels, and thus remain at risk for CHD. Accordingly, there is a need for additional LDL-C lowering agents.

The antisense compounds and methods provided herein are useful for the treatment of hypercholesterolemia. Treatment of hypercholesterolemia encompasses a therapeutic regimen that results in a clinically desirable outcome. For example, the antisense compounds and methods provided herein are useful for the treatment of elevated cholesterol, such as elevated LDL-C. In addition, the antisense compounds and methods provided herein may be used to reduce the risk of CHD, in individuals exhibiting one or more risk factors for CHD. Furthermore, the antisense compounds and methods provided herein may be used to treat and/or prevent atherosclerosis.

As illustrated herein, administration of an antisense oligonucleotide targeted to PCSK9 to animals fed a high-fat diet (an experimental model of hyperlipidemia) resulted in antisense inhibition of PCSK9, upregulation of the LDL-R, reduction of LDL-C levels, and reduction of liver triglycerides. Thus, it is demonstrated that in an experimental model of hyperlipidemia, antisense inhibition of PCSK9 results in lowering LDL-C levels. Accordingly, provided herein are methods for the treatment of reducing LDL-C levels through the administration of an antisense compound targeted to a PCSK9 nucleic acid. Increased LDL-C levels are considered a risk factor for CHD, and are also linked to atherosclerosis. Accordingly, also provided herein are methods for the reduction of CHD risk, and for the prevention and/or treatment of atherosclerosis. Also provided herein are methods for the treatment of conditions characterized by elevated liver triglycerides, such as hepatic steatosis.

In a particular embodiment, methods comprise the use of antisense compounds targeted to particularly advantageous sequences within a PCSK9 nucleic acid. The PCSK9 target sequences are selected on the basis of superior results shown in one or more selection criteria. The presently disclosed antisense compounds all exhibit high in vitro efficacy, determined using a battery of cell models to measure antisense regulation of PCSK9 expression. Further, preferred antisense compounds exhibit relatively high in vivo efficacy, determined using a transgenic mouse model that expresses human PCSK9 or a cynomolgus (“cyno”) monkey model. The antisense compound ISIS 405879 (SEQ ID NO: 248) displays superior results in suppressing PCSK9 mRNA expression in the liver of a host individual.

The present antisense compounds are tolerated to different extents when administered to a host individual. Increased tolerability can depend on a number of factors, including, but not limited to, the nucleotide sequence of the antisense compound, chemical modifications to the nucleotides, the particular motif of unmodified and modified nucleosides in the antisense compound, or combinations thereof. Antisense compounds that exhibit increased or superior tolerability are particularly preferred in the present methods. The present antisense compounds also can show different pharmacokinetic properties, depending on their nucleotide sequence, chemical modifications, motifs, or combinations thereof. Particularly preferred antisense compounds also exhibit favorable pharmacokinetics when administered to a host individual.

Certain Indications

In certain embodiments, the invention provides methods of treating an individual comprising administering one or more pharmaceutical compositions of the present invention. In certain embodiments, the individual has hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronary heart disease, a history of coronary heart disease, early onset coronary heart disease, acute coronary syndrome, one or more risk factors for coronary heart disease, type II diabetes, type II diabetes with dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia, hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, or non-alcoholic fatty liver disease.

Guidelines for lipid-lowering therapy were established in 2001 by Adult Treatment Panel III (ATP III) of the National Cholesterol Education Program (NCEP), and updated in 2004 (Grundy et al., Circulation, 2004, 110, 227-239). The guidelines include obtaining a complete lipoprotein profile, typically after a 9 to 12 hour fast, for determination of LDL-C, total cholesterol, and HDL-C levels. According to the most recently established guidelines, LDL-C levels of 130-159 mg/dL, 160-189 mg/dL, and greater than or equal to 190 mg/dL are considered borderline high, high, and very high, respectively. Total cholesterol levels of 200-239 and greater than or equal to 240 mg/dL are considered borderline high and high, respectively. HDL-C levels of less than 40 mg/dL are considered low.

In certain embodiments, the individual has been identified as in need of lipid-lowering therapy. In certain such embodiments, the individual has been identified as in need of lipid-lowering therapy according to the guidelines established in 2001 by Adult Treatment Panel III (ATP III) of the National Cholesterol Education Program (NCEP), and updated in 2004 (Grundy et al., Circulation, 2004, 110, 227-239). In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 190 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 160 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 130 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 100 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy should maintain LDL-C below 160 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy should maintain LDL-C below 130 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy should maintain LDL-C below 100 mg/dL. In certain such embodiments, the individual should maintain LDL-C below 70 mg/dL or even below 50 mg/dL.

In certain embodiments, the invention provides methods for reducing ApoB in an individual. In certain embodiments, the invention provides methods for reducing ApoB-containing lipoprotein in an individual. In certain embodiments, the invention provides methods for reducing LDL-C in an individual. In certain embodiments, the invention provides methods for reducing VLDL-C in an individual. In certain embodiments, the invention provides methods for reducing IDL-C in an individual. In certain embodiments, the invention provides methods for reducing non-HDL-C in an individual. In certain embodiments the invention provides methods for reducing Lp(a) in an individual. In certain embodiments, the invention provides methods for reducing serum triglyceride in an individual. In certain embodiments, the invention provides methods for reducing liver triglyceride in an individual. In certain embodiments, the invention provides methods for reducing Ox-LDL-C in an individual. In certain embodiments, the invention provides methods for reducing small LDL particles in an individual. In certain embodiments, the invention provides methods for reducing small VLDL particles in an individual. In certain embodiments, the invention provides methods for reducing phospholipids in an individual. In certain embodiments, the invention provides methods for reducing oxidized phospholipids in an individual.

In certain embodiments, the methods provided by the present invention do not lower HDL-C. In certain embodiments, the methods provided by the present invention do not result in accumulation of lipids in the liver.

In one embodiment are methods for decreasing LDL-C levels, or alternatively methods for treating hypercholesterolemia, by administering to an individual suffering from elevated LDL-C levels a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid. In another embodiment, a method of decreasing LDL-C levels comprises selecting an individual in need of a decrease in LDL-C levels, and administering to the individual a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid. In a further embodiment, a method of reducing coronary heart disease risk includes selecting an individual having elevated LDL-C levels and one or more additional indicators of coronary heart disease risk, and administering to the individual a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid.

In other embodiments, the LDL-C level is from 100-129 mg/dL, from 130 to 159 mg/dL, from 160-189 mg/dL, or greater than or equal to 190 mg/dL.

In one embodiment, administration of a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid is accompanied by monitoring of LDL-C levels in the serum of an individual, to determine an individual's response to administration of the antisense compound. An individual's response to administration of the antisense compound is used by a physician to determine the amount and duration of therapeutic intervention.

In one embodiment, administration of an antisense compound targeted to a PCSK9 nucleic acid results in LDL-C levels below 190 mg/dL, below 160 mg/dL, below 130 mg/dL, below 100 mg/dL, below 70 mg/dL, or below 50 mg/dL. In another embodiment, administration of an antisense compound targeted to a PCSK9 nucleic acid decreases LDL-C by at least 15%, by at least 25%, by at least 50%, by at least 60%, by at least 70%, by at least 75%, by at least 80%, by at least 85%, by at least 90%, or by at least 95%.

An individual having elevated LDL-C levels may also exhibit reduced HDL-C levels and/or elevated total cholesterol levels. Accordingly, in one embodiment a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid is administered to an individual having elevated LDL-C levels, who also has reduced HDL-C levels and/or elevated total cholesterol levels.

Individuals having elevated LDL-C levels may also exhibit elevated triglyceride levels. Accordingly, in one embodiment a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid is administered to an individual having elevated LDL-C levels, and also having elevated triglyceride levels.

Atherosclerosis can lead to coronary heart disease, stroke, or peripheral vascular disease. Elevated LDL-C levels are considered a risk factor in the development and progression of atherosclerosis. Accordingly, in one embodiment, a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid is administered to an individual having atherosclerosis. In a further embodiment, a therapeutically effective amount of antisense compound targeted to a PCSK9 nucleic acid is administered to an individual susceptible to atherosclerosis. Atherosclerosis is assessed directly through routine imaging techniques such as, for example, ultrasound imaging techniques that reveal carotid intimomedial thickness. Accordingly, treatment and/or prevention of atherosclerosis further include monitoring atherosclerosis through routine imaging techniques. In one embodiment, administration of an antisense compound targeted to a PCSK9 nucleic acid leads to a lessening of the severity of atherosclerosis, as indicated by, for example, a reduction of carotid intimomedial thickness in arteries.

Measurements of cholesterol, lipoproteins and triglycerides are obtained using serum or plasma collected from an individual. Methods of obtaining serum or plasma samples are routine, as are methods of preparation of the serum samples for analysis of cholesterol, triglycerides, and other serum markers.

A physician may determine the need for therapeutic intervention for individuals in cases where more or less aggressive LDL-lowering therapy is needed. The practice of the methods herein may be applied to any altered guidelines provided by the NCEP, or other entities that establish guidelines for physicians used in treating any of the diseases or conditions listed herein, for determining coronary heart disease risk and diagnosing metabolic syndrome.

In one embodiment, administration of an antisense compound targeted to a PCSK9 nucleic acid is parenteral administration. Parenteral administration may be intravenous or subcutaneous administration. Accordingly, in another embodiment, administration of an antisense compound targeted to a PCSK9 nucleic acid is intravenous or subcutaneous administration. Administration may include multiple doses of an antisense compound targeted to a PCSK9 nucleic acid.

In certain embodiments, a pharmaceutical composition comprising an antisense compound targeted to PCSK9 is for use in therapy. In certain embodiments, the therapy is the reduction of LDL-C, ApoB, VLDL-C, IDL-C, non-HDL-C, Lp(a), serum triglyceride, liver triglyceride, Ox-LDL-C, small LDL particles, small VLDL, phospholipids, or oxidized phospholipids in an individual. In certain embodiments, the therapy is the treatment of hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronary heart disease, acute coronary syndrome, a history of coronary heart disease, early onset coronary heart disease, one or more risk factors for coronary heart disease, type II diabetes, type II diabetes with dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia, hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, or non-alcoholic fatty liver disease. In additional embodiments, the therapy is the reduction of CHD risk. In certain aspects, the therapy is prevention of atherosclerosis. In certain embodiments, the therapy is the prevention of coronary heart disease.

In certain embodiments, a pharmaceutical composition comprising an antisense compound targeted to PCSK9 is used for the preparation of a medicament for reducing LDL-C, ApoB, VLDL-C, IDL-C, non-HDL-C, Lp(a), serum triglyceride, liver triglyceride, Ox-LDL-C, small LDL particles, small VLDL, phospholipids, or oxidized phospholipids in an individual. In certain embodiments pharmaceutical composition comprising an antisense compound targeted to PCKS9 is used for the preparation of a medicament for reducing coronary heart disease risk. In certain embodiments, an antisense compound targeted to PCSK9 is used for the preparation of a medicament for the treatment of hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronary heart disease, a history of coronary heart disease, early onset coronary heart disease, one or more risk factors for coronary heart disease, type II diabetes, type II diabetes with dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia, hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, or non-alcoholic fatty liver disease.

Certain Combination Therapies

In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with one or more other pharmaceutical agents. In certain embodiments, such one or more other pharmaceutical agents are designed to treat the same disease or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat a different disease or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat an undesired effect of one or more pharmaceutical compositions of the present invention. In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with another pharmaceutical agent to treat an undesired effect of that other pharmaceutical agent. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at the same time. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at different times. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared together in a single formulation. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared separately. For example, a composition may comprise a pharmaceutical agent for separate, sequential, or simultaneous administration with an antisense compound.

In certain embodiments, pharmaceutical agents that may be co-administered with a pharmaceutical composition of the present invention include lipid-lowering agents or LXR agonists. In certain such embodiments, pharmaceutical agents that may be co-administered with a pharmaceutical composition of the present invention include, but are not limited to atorvastatin, simvastatin, rosuvastatin, and ezetimibe. In certain such embodiments, the lipid-lowering agent is administered prior to administration of a pharmaceutical composition of the present invention. In certain such embodiments, the lipid-lowering agent is administered following administration of a pharmaceutical composition of the present invention. In certain such embodiments the lipid-lowering agent is administered at the same time as a pharmaceutical composition of the present invention. In certain such embodiments the dose of a co-administered lipid-lowering agent is the same as the dose that would be administered if the lipid-lowering agent was administered alone. In certain such embodiments the dose of a co-administered lipid-lowering agent is lower than the dose that would be administered if the lipid-lowering agent was administered alone. In certain such embodiments the dose of a co-administered lipid-lowering agent is greater than the dose that would be administered if the lipid-lowering agent was administered alone.

In certain embodiments, a co-administered lipid-lowering agent is a HMG-CoA reductase inhibitor. In certain such embodiments the HMG-CoA reductase inhibitor is a statin. In certain such embodiments, the statin is selected from, for example, atorvastatin, simvastatin, pravastatin, fluvastatin, and rosuvastatin.

In certain embodiments, a co-administered lipid-lowering agent is a cholesterol absorption inhibitor. In certain such embodiments, cholesterol absorption inhibitor is ezetimibe.

In certain embodiments, a co-administered lipid-lowering agent is a co-formulated HMG-CoA reductase inhibitor and cholesterol absorption inhibitor. In certain such embodiments the co-formulated lipid-lowering agent is ezetimibe/simvastatin.

In certain embodiments, a co-administered lipid-lowering agent is a microsomal triglyceride transfer protein inhibitor (MTP inhibitor).

In certain embodiments, a co-administered lipid-lowering agent is an oligonucleotide targeted to ApoB.

In certain embodiments, a co-administered pharmaceutical agent is a bile acid sequestrant. In certain such embodiments, the bile acid sequestrant is selected from cholestyramine, colestipol, and colesevelam.

In certain embodiments, a co-administered pharmaceutical agent is a nicotinic acid. In certain such embodiments, the nicotinic acid is selected from immediate release nicotinic acid, extended release nicotinic acid, and sustained release nicotinic acid.

In certain embodiments, a co-administered pharmaceutical agent is a fibric acid. In certain such embodiments, a fibric acid is selected from gemfibrozil, fenofibrate, clofibrate, bezafibrate, and ciprofibrate.

Further examples of pharmaceutical agents that may be co-administered with a pharmaceutical composition of the present invention include, but are not limited to, corticosteroids, including but not limited to prednisone; LXR agonists; immunoglobulins, including, but not limited to intravenous immunoglobulin (IVIg); analgesics (e.g., acetaminophen); anti-inflammatory agents, including, but not limited to non-steroidal anti-inflammatory drugs (e.g., ibuprofen, COX-1 inhibitors, and COX-2, inhibitors); salicylates; antibiotics; antivirals; antifungal agents; antidiabetic agents (e.g., biguanides, glucosidase inhibitors, insulins, sulfonylureas, and thiazolidenediones); adrenergic modifiers; diuretics; hormones (e.g., anabolic steroids, androgen, estrogen, calcitonin, progestin, somatostan, and thyroid hormones); immunomodulators; muscle relaxants; antihistamines; osteoporosis agents (e.g., biphosphonates, calcitonin, and estrogens); prostaglandins, antineoplastic agents; psychotherapeutic agents; sedatives; poison oak or poison sumac products; antibodies; and vaccines.

In certain embodiments, the pharmaceutical compositions of the present invention may be administered in conjunction with a lipid-lowering therapy. In certain such embodiments, a lipid-lowering therapy is therapeutic lifestyle change. In certain such embodiments, a lipid-lowering therapy is LDL apheresis.

Antisense Compounds

Oligomeric compounds include, but are not limited to, oligonucleotides, oligonucleosides, oligonucleotide analogs, oligonucleotide mimetics, antisense compounds, antisense oligonucleotides, and siRNAs. An oligomeric compound may be “antisense” to a target nucleic acid, meaning that is capable of undergoing hybridization to a target nucleic acid through hydrogen bonding.

In certain embodiments, an antisense compound has a nucleobase sequence that, when written in the 5′ to 3′ direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is targeted. In certain such embodiments, an antisense oligonucleotide has a nucleobase sequence that, when written in the 5′ to 3′ direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is targeted.

In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid is 8 to 80, 12 to 50, 12 to 30 or 15 to 30 subunits in length. In other words, antisense compounds are from 8 to 80, 12 to 50, 12 to 30 or 15 to 30 linked subunits. In certain such embodiments, the antisense compounds are 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or subunits in length.

In certain embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 12 to 30 nucleotides in length. In certain such embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.

In certain embodiment, an antisense compound targeted to a PCSK9 nucleic acid is 15 to 30 subunits in length. In other words, antisense compounds are from 15 to 30 linked subunits. In certain such embodiments, the antisense compounds are 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 subunits in length.

In certain embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 15 to 30 nucleotides in length. In certain such embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.

In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid is 18 to 24 subunits in length. In other words, antisense compounds are from 18 to 24 linked subunits. In one embodiment, the antisense compounds are 18, 19, 20, 21, 22, 23, or 24 subunits in length.

In certain embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 18 to 24 nucleotides in length. In certain such embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 18, 19, 20, 21, 22, 23, or 24 nucleotides in length.

In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid is 19 to 22 subunits in length. In other words, antisense compounds are from 19 to 22 linked subunits. This embodies antisense compounds of 19, 20, 21, or 22 subunits in length.

In certain embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 19 to 22 nucleotides in length. In certain such embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 19, 20, 21, or 22 nucleotides in length.

In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid is 20 subunits in length. In certain such embodiments, antisense compounds are 20 linked subunits in length.

In certain embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 20 nucleotides in length. In certain such embodiments, an antisense oligonucleotide targeted to an PCSK9 nucleic acid is 20 linked nucleotides in length.

In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common.

In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence target a region identified herein.

In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid may target the following nucleotide regions of SEQ ID NO: 1: 294-317, 406-440, 406-526, 410-436, 410-499, 446-526, 545-581, 591-619, 591-704, 591-743, 595-622, 600-626, 600-639, 600-670, 601-628, 602-628, 603-630, 611-636, 620-647, 638-665, 648-674, 657-684, 705-743, 782-810, 821-859, 835-859, 835-917, 835-942, 860-887, 860-899, 860-909, 860-917, 869-895, 878-905, 888-909, 923-952, 960-1034, 960-1173, 960-986, 967-991, 970-1023, 970-1064, 970-1117, 970-996, 977-1004, 985-1011, 989-1016, 992-1019, 997-1024, 997-1024, 998-1025, 999-1026, 1000-1027, 1001-1028, 1002-1029, 1003-1029, 1004-1029, 1005-1029, 1006-1029, 1007-1034, 1036-1061, 1045-1072, 1076-1096, 1088-1115, 1098-1123, 1200-1251, 1210-1237, 1219-1245, 1228-1251, 1273-1444, 1295-1316, 1318-1345, 1328-1354, 1337-1361, 1344-1371, 1354-1377, 1380-1406, 1389-1416, 1400-1426, 1409-1434, 1465-1491, 1465-1602, 1474-1499, 1482-1519, 1513-1540, 1523-1549, 1526-1602, 1526-1624, 1532-1558, 1541-1568, 1552-1579, 1560-1587, 1561-1589, 1564-1591, 1565-1592, 1566-1592, 1567-1592, 1570-1597, 1571-1599, 1605-1706, 1628-1706, 1640-1666, 1672-1698, 1681-1706, 1735-1761, 1735-1765, 1740-1765, 1849-1876, 1849-1879, 1850-1877, 1851-1877, 1852-1878, 1852-1879, 1853-1879, 1854-1879, 1905-1955, 1915-1942, 1916-1943, 1917-1944, 1918-1945, 1919-1946, 1920-1939, 1920-1947, 1921-1948, 1922-1949, 1923-1950, 1924-1951, 1925-1952, 1926-1952, 1927-1952, 1928-1955, 1962-2059, 2040-2126, 2100-2126, 2100-2139, 2100-2206, 2101-2126, 2305-2332, 2305-2354, 2306-2333, 2307-2334, 2308-2334, 2309-2334, 2310-2334, 2410-2434, 2504-2528, 2509-2528, 2582-2625, 2606-2668, 2828-2855, 2832-2851, 2900-2927, 2900-2929, 2902-2927, 2983-3007, 2983-3013, 3227-3252, 3227-3456, 3472-3496, or 3543-3569.

In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 1: 294-317, 406-440, 406-526, 410-436, 410-499, 446-526, 545-581, 591-619, 591-704, 591-743, 595-622, 600-626, 600-639, 600-670, 601-628, 602-628, 603-630, 611-636, 620-647, 638-665, 648-674, 657-684, 705-743, 782-810, 821-859, 835-859, 835-917, 835-942, 860-887, 860-899, 860-909, 860-917, 869-895, 878-905, 888-909, 923-952, 960-1034, 960-1173, 960-986, 967-991, 970-1023, 970-1064, 970-1117, 970-996, 977-1004, 985-1011, 989-1016, 992-1019, 997-1024, 997-1024, 998-1025, 999-1026, 1000-1027, 1001-1028, 1002-1021, 1002-1029, 1003-1029, 1004-1029, 1005-1029, 1006-1029, 1007-1034, 1036-1061, 1045-1072, 1076-1096, 1088-1115, 1098-1123, 1200-1251, 1210-1237, 1219-1245, 1228-1251, 1273-1444, 1295-1316, 1318-1345, 1328-1354, 1337-1361, 1344-1371, 1354-1377, 1380-1406, 1389-1416, 1400-1426, 1409-1434, 1465-1491, 1465-1602, 1474-1499, 1482-1519, 1513-1540, 1523-1549, 1526-1602, 1526-1624, 1532-1558, 1541-1568, 1552-1579, 1560-1587, 1561-1589, 1564-1591, 1565-1592, 1566-1592, 1567-1592, 1570-1597, 1571-1599, 1605-1706, 1628-1706, 1640-1666, 1672-1698, 1681-1706, 1735-1761, 1735-1765, 1740-1765, 1849-1876, 1849-1879, 1850-1877, 1851-1877, 1852-1878, 1852-1879, 1853-1879, 1854-1879, 1905-1955, 1915-1942, 1916-1943, 1917-1944, 1918-1945, 1919-1946, 1920-1939, 1920-1947, 1921-1948, 1922-1949, 1923-1950, 1924-1951, 1925-1952, 1926-1952, 1927-1952, 1928-1955, 1962-2059, 2040-2126, 2100-2126, 2100-2139, 2100-2206, 2101-2126, 2305-2332, 2305-2354, 2306-2333, 2307-2334, 2308-2334, 2309-2334, 2310-2334, 2410-2434, 2504-2528, 2509-2528, 2582-2625, 2606-2668, 2828-2855, 2832-2851, 2900-2927, 2900-2929, 2902-2927, 2983-3007, 2983-3013, 3227-3252, 3227-3456, 3472-3496, or 3543-3569.

In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid may target the following nucleotide regions of SEQ ID NO: 2: 2274-2400, 2274-2575, 2433-2570, 2433-2579, 2549-2575, 2552-2579, 2585-2638, 2605-2638, 3056-3075, 4150-5159, 4306-4325, 5590-5618, 5667-5686, 6444-6463, 6482-6518, 6492-6518, 6528-6555, 6528-6623, 6534-6561, 6535-6562, 6536-6563, 6537-6563, 6538-6565, 6539-6565, 6540-6567, 6541-6567, 6542-6569, 6546-6573, 6557-6584, 6575-6602, 6585-6611, 6594-6621, 6596-6623, 6652-6671, 7099-7118, 7556-7584, 8836-8855, 8948-8967, 9099-9118, 9099-9168, 9130-9168, 9207-9233, 9207-9235, 9209-9235, 10252-10271, 10633-10652, 11308-11491, 12715-12734, 12928-12947, 13681-13700, 13746-13779, 13816-13847, 13903-13945, 13977-14141, 14179-14198, 14267-14286, 14397-14423, 14441-14460, 14494-14513, 14494-14543, 14524-14543, 14601-14650, 14670-14700, 14675-14700, 14801-14828, 14877-14912, 14877-14915, 14877-14973, 14916-14943, 14916-14973, 14925-14951, 14934-14963, 14946-14973, 14979-14998, 15254-15280, 15254-15328, 15264-15290, 15279-15305, 15291-15318, 15292-15319, 15293-15320, 15294-15321, 15294-15321, 15295-15322, 15296-15323, 15297-15323, 15298-15323, 15299-15323, 15300-15323, 15301-15328, 15330-15355, 15330-15490, 15339-15366, 15358-15490, 16134-16153, 16668-16687, 17267-17286, 18377-18427, 18561-18580, 18591-18618, 18591-18646, 18591-18668, 18695-18746, 18705-18730, 18709-18736, 18719-18746, 19203-20080, 19931-19952, 19954-19981, 19964-19990, 19973-19999, 19982-20009, 19992-20016, 20016-20042, 20025-20052, 20036-20062, 20045-20070, 20100-20119, 20188-20207, 20624-20650, 20624-20759, 20629-20804, 20633-20660, 20635-20781, 20643-20662, 20657-20676, 20670-20697, 20680-20706, 20683-20781, 20689-20715, 20698-20725, 20709-20736, 20717-20744, 20718-20745, 20719-20746, 20720-20747, 20721-20748, 20722-20749, 20727-20752, 20735-20759, 20762-21014, 20785-21014, 21082-21107, 21082-21152, 21091-21114, 21118-21144, 21127-21152, 21181-21209, 21181-21211, 21183-21211, 21481-21500, 21589-21608, 21692-21719, 22000-22227, 22096-22115, 22096-22223, 22096-22311, 22133-22160, 22133-22163, 22134-22161, 22135-22162, 22136-22163, 22137-22163, 22138-22163, 22189-22239, 22199-22226, 22199-22227, 22200-22227, 22201-22228, 22202-22229, 22203-22230, 22204-22231, 22205-22232, 22206-22233, 22207-22234, 22208-22235, 22209-22236, 22210-22236, 22210-22239, 22211-22236, 22212-22239, 22292-22311, 23985-24054, 24035-24134, 24095-24121, 24858-24877, 24907-24926, 25413-25432, 25994-26013, 26112-26139, 26112-26161, 26112-27303, 26113-26140, 26114-26141, 26115-26141, 26116-26141, 26117-26141, 26117-26475, 26118-26141, 26120-26141, 26132-26151, 26142-26161, 26217-26241, 26311-26335, 26389-26432, 26456-26576, 26635-26662, 26707-26734, 26707-26736, 26790-26820, 27034-27263, 27279-27303, or 27350-27376.

In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 2: 2274-2400, 2274-2575, 2433-2570, 2433-2579, 2549-2575, 2552-2579, 2585-2638, 2605-2638, 3056-3075, 4150-5159, 4306-4325, 5590-5618, 5667-5686, 6444-6463, 6482-6518, 6492-6518, 6528-6555, 6528-6623, 6534-6561, 6535-6562, 6536-6563, 6537-6563, 6538-6565, 6539-6565, 6540-6567, 6541-6567, 6542-6569, 6546-6573, 6557-6584, 6575-6602, 6585-6611, 6594-6621, 6596-6623, 6652-6671, 7099-7118, 7556-7584, 8836-8855, 8948-8967, 9099-9118, 9099-9168, 9130-9168, 9207-9233, 9207-9235, 9209-9235, 10252-10271, 10633-10652, 11308-11491, 12715-12734, 12928-12947, 13681-13700, 13746-13779, 13816-13847, 13903-13945, 13977-14141, 14179-14198, 14267-14286, 14397-14423, 14441-14460, 14494-14513, 14494-14543, 14524-14543, 14601-14650, 14670-14700, 14675-14700, 14801-14828, 14877-14912, 14877-14915, 14877-14973, 14916-14943, 14916-14973, 14925-14951, 14934-14963, 14946-14973, 14979-14998, 15254-15280, 15254-15328, 15264-15290, 15279-15305, 15291-15318, 15292-15319, 15293-15320, 15294-15321, 15294-15321, 15295-15322, 15296-15323, 15297-15323, 15298-15323, 15299-15323, 15300-15323, 15301-15328, 15330-15355, 15330-15490, 15339-15366, 15358-15490, 16134-16153, 16668-16687, 17267-17286, 18377-18427, 18561-18580, 18591-18618, 18591-18646, 18591-18668, 18695-18746, 18705-18730, 18709-18736, 18719-18746, 19203-20080, 19931-19952, 19954-19981, 19964-19990, 19973-19999, 19982-20009, 19992-20016, 20016-20042, 20025-20052, 20036-20062, 20045-20070, 20100-20119, 20188-20207, 20624-20650, 20624-20759, 20629-20804, 20633-20660, 20635-20781, 20643-20662, 20657-20676, 20670-20697, 20680-20706, 20683-20781, 20689-20715, 20698-20725, 20709-20736, 20717-20744, 20718-20745, 20719-20746, 20720-20747, 20721-20748, 20722-20749, 20727-20752, 20735-20759, 20762-21014, 20785-21014, 21082-21107, 21082-21152, 21091-21114, 21118-21144, 21127-21152, 21181-21209, 21181-21211, 21183-21211, 21481-21500, 21589-21608, 21692-21719, 22000-22227, 22096-22115, 22096-22223, 22096-22311, 22133-22160, 22133-22163, 22134-22161, 22135-22162, 22136-22163, 22137-22163, 22138-22163, 22189-22239, 22199-22226, 22199-22227, 22200-22227, 22201-22228, 22202-22229, 22203-22230, 22204-22231, 22205-22232, 22206-22233, 22207-22234, 22208-22235, 22209-22236, 22210-22236, 22210-22239, 22211-22236, 22212-22239, 22292-22311, 23985-24054, 24035-24134, 24095-24121, 24858-24877, 24907-24926, 25413-25432, 25994-26013, 26112-26139, 26112-26161, 26112-27303, 26113-26140, 26114-26141, 26115-26141, 26116-26141, 26117-26141, 26117-26475, 26118-26141, 26120-26141, 26132-26151, 26142-26161, 26217-26241, 26311-26335, 26389-26432, 26456-26576, 26635-26662, 26707-26734, 26707-26736, 26790-26820, 27034-27263, 27279-27303, or 27350-27376.

In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid may target the following nucleotide regions of SEQ ID NO: 3: 220-253, 290-321, 377-419, 451-615, 653-672, 741-760, 871-897, 915-934, 968-1017, 1075-1124, 1075-1174, 1144-1174, 1275-1302, 1315-1341, 1351-1389, 1351-1447, 1365-1439, 1390-1417, 1390-1429, 1390-1439, 1399-1425, 1408-1435, 1420-1447, 1453-1482, 1490-1516, 1490-1564, 1500-1526, 1515-1541, 1527-1553, 1527-1554, 1528-1554, 1529-1555, 1529-1556, 1530-1556, 1530-1557, 1531-1557, 1532-1558, 1533-1559, 1534-1559, 1535-1559, 1536-1559, 1537-1564, 1566-1602, 1566-1681, 1606-1626, 1618-1645, 1626-1653, 1684-1703, 1730-1781, 1740-1767, 1749-1775, 1758-1781, 1820-1847, 1820-1877, 1822-2198, 1830-1856, 1839-1865, 1840-1867, 1898-1924, 1898-2035, 1903-2127, 1907-1934, 1911-1938, 1946-1971, 1954-1980, 1959-2035, 1959-2057, 1963-1988, 1967-2035, 1972-1999, 1982-2008, 1991-2018, 1993-2019, 1995-2022, 1996-2023, 1997-2024, 1998-2025, 1999-2025, 2000-2025, 2009-2035, 2038-2139, 2061-2139, 2073-2099, 2078-2104, 2105-2131, 2112-2139, 2168-2198, 2170-2177, 2245-2284, 2295-2394, 2355-2381, 2355-2394, 2405-2461, 2560-2587, 2560-2609, 2561-2588, 2562-2589, 2563-2589, 2564-2589, 2565-2589, 2566-2589, 2567-2589, 2568-2589, 2665-2689, 2759-2783, 2837-2880, 2904-2923, 3005-3024, 3005-3174, 3083-3110, 3155-3184, 3238-3268, 3482-3711, 3727-3751, or 3798-3824.

In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 3: 220-253, 290-321, 377-419, 451-615, 653-672, 741-760, 871-897, 915-934, 968-1017, 1075-1124, 1075-1174, 1144-1174, 1275-1302, 1315-1341, 1351-1389, 1351-1447, 1365-1439, 1390-1417, 1390-1429, 1390-1439, 1399-1425, 1408-1435, 1420-1447, 1453-1482, 1490-1516, 1490-1564, 1500-1526, 1515-1541, 1527-1553, 1527-1554, 1528-1554, 1529-1555, 1529-1556, 1530-1556, 1530-1557, 1531-1557, 1532-1558, 1533-1559, 1534-1559, 1535-1559, 1536-1559, 1537-1564, 1566-1602, 1566-1681, 1606-1626, 1618-1645, 1626-1653, 1684-1703, 1730-1781, 1740-1767, 1749-1775, 1758-1781, 1820-1847, 1820-1877, 1822-2198, 1830-1856, 1839-1865, 1840-1867, 1898-1924, 1898-2035, 1903-2127, 1907-1934, 1911-1938, 1946-1971, 1954-1980, 1959-2035, 1959-2057, 1963-1988, 1967-2035, 1972-1999, 1982-2008, 1991-2018, 1993-2019, 1995-2022, 1996-2023, 1997-2024, 1998-2025, 1999-2025, 2000-2025, 2009-2035, 2038-2139, 2061-2139, 2073-2099, 2078-2104, 2105-2131, 2112-2139, 2168-2198, 2170-2177, 2245-2284, 2295-2394, 2355-2381, 2355-2394, 2405-2461, 2560-2587, 2560-2609, 2561-2588, 2562-2589, 2563-2589, 2564-2589, 2565-2589, 2566-2589, 2567-2589, 2568-2589, 2665-2689, 2759-2783, 2837-2880, 2904-2923, 3005-3024, 3005-3174, 3083-3110, 3155-3184, 3238-3268, 3482-3711, 3727-3751, or 3798-3824.

In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid may target the following nucleotide regions of SEQ ID NO: 1: 320-405, 441-445, 527-544, 582-590, 744-781, 811-820, 918-922, 953-959, 1034-1036, 1152-1153, 1174-1199, 1251-1272, 1445-1464, 1603-1604, 1625-1627, 1707-1734, 1766-1811, 1832-1848, 1880-1904, 1956-1961, 1982-1939, 2030-2039, 2060-2099, 2140-2149, 2170-2186, 2207-2304, 2355-2409, 2435-2503, 2529-2581, 2626-2648, 2669-2749, 2770-2827, 2856-2876, 2891-2899, 2930-2982, 3014-3226, 3253-3436, 3457-3471, or 3497-3542.

In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common.

In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 1: 320-405, 441-445, 527-544, 582-590, 744-781, 811-820, 918-922, 953-959, 1034-1036, 1152-1153, 1174-1199, 1251-1272, 1445-1464, 1603-1604, 1625-1627, 1707-1734, 1766-1811, 1832-1848, 1880-1904, 1956-1961, 1982-1939, 2030-2039, 2060-2099, 2140-2149, 2170-2186, 2207-2304, 2355-2409, 2435-2503, 2529-2581, 2626-2648, 2669-2749, 2770-2827, 2856-2876, 2891-2899, 2930-2982, 3014-3226, 3253-3436, 3457-3471, or 3497-3542.

In certain embodiments, a shortened or truncated antisense compound targeted to a PCSK9 nucleic acid has a single subunit deleted from the 5′ end (5′ truncation), or alternatively from the 3′ end (3′ truncation). A shortened or truncated antisense compound targeted to a PCSK9 nucleic acid may have two subunits deleted from the 5′ end, or alternatively may have two subunits deleted from the 3′ end, of the antisense compound. Alternatively, the deleted nucleosides may be dispersed throughout the antisense compound, for example, in an antisense compound having one nucleoside deleted from the 5′ end and one nucleoside deleted from the 3′ end.

When a single additional subunit is present in a lengthened antisense compound, the additional subunit may be located at the 5′ or 3′ end of the antisense compound. When two are more additional subunits are present, the added subunits may be adjacent to each other, for example, in an antisense compound having two subunits added to the 5′ end (5′ addition), or alternatively to the 3′ end (3′ addition), of the antisense compound. Alternatively, the added subunits may be dispersed throughout the antisense compound, for example, in an antisense compound having one subunit added to the 5′ end and one subunit added to the 3′ end.

It is possible to increase or decrease the length of an antisense compound, such as an antisense oligonucleotide, and/or introduce mismatch bases without eliminating activity. For example, in Woolf et al. (Proc. Natl. Acad. Sci. USA 89:7305-7309, 1992), a series of antisense oligonucleotides 13-25 nucleobases in length were tested for their ability to induce cleavage of a target RNA in an oocyte injection model. Antisense oligonucleotides 25 nucleobases in length with 8 or 11 mismatch bases near the ends of the antisense oligonucleotides were able to direct specific cleavage of the target mRNA, albeit to a lesser extent than the antisense oligonucleotides that contained no mismatches. Similarly, target specific cleavage was achieved using 13 nucleobase antisense oligonucleotides, including those with 1 or 3 mismatches.

Gautschi et al (J. Natl. Cancer Inst. 93:463-471, March 2001) demonstrated the ability of an oligonucleotide having 100% complementarity to the bcl-2 mRNA and having 3 mismatches to the bcl-xL mRNA to reduce the expression of both bcl-2 and bcl-xL in vitro and in vivo. Furthermore, this oligonucleotide demonstrated potent anti-tumor activity in vivo.

Maher and Dolnick (Nuc. Acid. Res. 16:3341-3358, 1988) tested a series of tandem 14 nucleobase antisense oligonucleotides, and a 28 and 42 nucleobase antisense oligonucleotides comprised of the sequence of two or three of the tandem antisense oligonucleotides, respectively, for their ability to arrest translation of human DHFR in a rabbit reticulocyte assay. Each of the three 14 nucleobase antisense oligonucleotides alone was able to inhibit translation, albeit at a more modest level than the 28 or 42 nucleobase antisense oligonucleotides.

PCT/US2007/068404 describes incorporation of chemically-modified high-affinity nucleotides into short antisense compounds about 8-16 nucleobases in length and that such compounds are useful in the reduction of target RNAs in animals with increased potency and improved therapeutic index.

In certain embodiments, antisense compounds targeted to a PCSK9 nucleic acid are short antisense compounds. In certain embodiments, such short antisense compounds are oligonucleotide compounds. In certain embodiments, such short antisense compounds are about 8 to 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 nucleotides in length and comprises a gap region flanked on each side by a wing, wherein each wing independently consists of 1 to 3 nucleotides. Preferred motifs include but are not limited to wing-deoxy gap-wing motifs selected from 3-10-3, 2-10-3, 2-10-2, 1-10-1, 2-8-2, 1-8-1, 3-6-3 or 1-6-1.

Antisense compounds targeted to a PCSK9 nucleic acid are synthesized in vitro and do not include antisense compositions of biological origin, or genetic vector constructs designed to direct the in vivo synthesis of antisense molecules.

In certain embodiments, an antisense compound is targeted to a region of a PCSK9 nucleic acid that does not contain a single nucleotide polymorphism (SNPs). In certain embodiments, an antisense compound is targeted to a region of a PCSK9 nucleic acid that does contain a single nucleotide polymorph (SNPs). A single nucleotide polymorphism refers to polymorphisms that are the result of a single nucleotide alteration or the existence of two or more alternative sequences which can be, for example, different allelic forms of a gene. A polymorphism may comprise one or more base changes including, for example, an insertion, a repeat, or a deletion. In certain embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid overlaps with a SNP at the following positions: 428, 432, 449, 996, 1011, 1044, 1317, 1565, 1617, 1618, 1671, 1711, 1722, 1836, 1911. In certain embodiments, the compounds provided herein that target a region of a PCSK9 nucleic acid that contains one or more SNPs will contain the appropriate base substitution, insertion, repeat or deletion such that the compound is fully complementary to the altered PCSK9 nucleic acid sequence.

A subgroup of these antisense compounds were selected for further characterization, based on the ability of the selected antisense compounds to inhibit PCSK9 expression selectively and effectively in cell culture assays, set forth in the Examples. The antisense compounds selected for further evaluation are listed in Table 1. The 5′ and 3′ boundaries of the PCSK9 target sequence are shown for each antisense compound, with reference to the nucleotide positions of SEQ ID NO: 1.

“Motif” in Table 1 refers to the use of chemically modified nucleosides at the 5′ and 3′ ends of the antisense compounds. In a “5-10-5” motif, for example, five chemical modified nucleosides at both the 5′ and 3′ ends flank ten unmodified nucleosides in the center of the antisense compound. As disclosed in more detail below, modified nucleosides may make the antisense compounds more resistant to nucleases, among other things, which generally improves the pharmacodynamic and pharmacokinetic properties of the antisense compounds when administered to an individual.

Table 1 further discloses the nucleobase sequence of the antisense compounds. In certain embodiments, an antisense compound has a nucleobase sequence that, when written in the 5′ to 3′ direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is targeted.

TABLE 1 5′ Target 3′ Target SEQ ID Isis No. Site Site Sequence 5′-3′ Motif NO 405881 1005 1024 CACCCTTGGCCACGCCGGCA 5-10-5 250 399819 2509 2528 CCCACTCAAGGGCCAGGCCA 5-10-5 65 395165 1004 1023 ACCCTTGGCCACGCCGGCAT 5-10-5 28 405879 1002 1021 CCTTGGCCACGCCGGCATCC 5-10-5 248 406008 602 621 CTTGGTGAGGTATCCCCGGC 5-10-5 188 405891 1567 1586 TCCTCAGGGAACCAGGCCTC 5-10-5 352 395186 2100 2119 CTTTGCATTCCAGACCTGGG 5-10-5 60 405988 1854 1873 GGCAGCACCTGGCAATGGCG 5-10-5 381 405994 1928 1947 GCAGTGGACACGGGTCCCCA 5-10-5 400 406023 787 806 TGGTATTCATCCGCCCGGTA 5-10-5 212 395187 2310 2329 GGCAGCAGATGGCAACGGCT 5-10-5 62 395185 1920 1939 CACGGGTCCCCATGCTGGCC 5-10-5 59 406033 967 986 CCTGCCAGGTGGGTGCCATG 5-10-5 237 405923 1295 1314 GGCATTGGTGGCCCCAACTG 5-10-5 288 399900 1569 1588 GGTCCTCAGGGAACCAGGCC 3-14-3 50 405995 1930 1949 TGGCAGTGGACACGGGTCCC 5-10-5 402 405991 1922 1941 GACACGGGTCCCCATGCTGG 5-10-5 394 406005 559 578 CGGGCAGTGCGCTCTGACTG 5-10-5 180 399793 417 436 CCTCGGAACGCAAGGCTAGC 5-10-5 8 395152 410 429 ACGCAAGGCTAGCACCAGCT 5-10-5 7 Antisense Compound Motifs

In certain embodiments, antisense compounds targeted to a PCSK9 nucleic acid have chemically modified subunits arranged in patterns, or motifs, to confer to the antisense compounds properties such as enhanced the inhibitory activity, increased binding affinity for a target nucleic acid, or resistance to degradation by in vivo nucleases.

Chimeric antisense compounds typically contain at least one region modified so as to confer increased resistance to nuclease degradation, increased cellular uptake, increased binding affinity for the target nucleic acid, and/or increased inhibitory activity. A second region of a chimeric antisense compound may serve as a substrate for the cellular endonuclease RNase H, which cleaves the RNA strand of an RNA:DNA duplex.

Antisense compounds having a gapmer motif are considered chimeric antisense compounds. In a gapmer an internal position having a plurality of nucleotides that supports RNaseH cleavage is positioned between external regions having a plurality of nucleotides that are chemically distinct from the nucleosides of the internal region. In the case of an antisense oligonucleotide having a gapmer motif, the gap segment generally serves as the substrate for endonuclease cleavage, while the wing segments comprise modified nucleosides. The regions of a gapmer are differentiated by the types of sugar moieties comprising each distinct region. The types of sugar moieties that are used to differentiate the regions of a gapmer may in some embodiments include β-D-ribonucleosides, β-D-deoxyribonucleosides, 2′-modified nucleosides (such 2′-modified nucleosides may include 2′-MOE, and 2′-O—CH₃, among others), and bicyclic sugar modified nucleosides (such bicyclic sugar modified nucleosides may include those having a 4′-(CH2)n-O-2′ bridge, where n=1 or n=2). In general, each distinct region comprises uniform sugar moieties. The wing-gap-wing motif is frequently described as “X-Y-Z”, where “X” represents the length of the 5′ wing region, “Y” represents the length of the gap region, and “Z” represents the length of the 3′ wing region.

In some embodiments, an antisense compound targeted to a PCSK9 nucleic acid has a gap-widened motif. In other embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid has a gap-widened motif.

The gap-widened antisense oligonucleotides described herein may have various wing-gap-wing motifs selected from: 1-16-1, 2-15-1, 1-15-2, 1-14-3, 3-14-1, 2-14-2, 1-13-4, 4-13-1, 2-13-3, 3-13-2, 1-12-5, 5-12-1, 2-12-4, 4-12-2, 3-12-3, 1-11-6, 6-11-1, 2-11-5, 5-11-2, 3-11-4, 4-11-3, 1-17-1, 2-16-1, 1-16-2, 1-15-3, 3-15-1, 2-15-2, 1-14-4, 4-14-1, 2-14-3, 3-14-2, 1-13-5, 5-13-1, 2-13-4, 4-13-2, 3-13-3, 1-12-6, 6-12-1, 2-12-5, 5-12-2, 3-12-4, 4-12-3, 1-11-7, 7-11-1, 2-11-6, 6-11-2, 3-11-5, 5-11-3, 4-11-4, 1-18-1, 1-17-2, 2-17-1, 1-16-3, 1-16-3, 2-16-2, 1-15-4, 4-15-1, 2-15-3, 3-15-2, 1-14-5, 5-14-1, 2-14-4, 4-14-2, 3-14-3, 1-13-6, 6-13-1, 2-13-5, 5-13-2, 3-13-4, 4-13-3, 1-12-7, 7-12-1, 2-12-6, 6-12-2, 3-12-5, 5-12-3, 4-12-4, 1-11-8, 8-11-1, 2-11-7, 7-11-2, 3-11-6, 6-11-3, 4-11-5, 5-11-4, 1-18-1, 1-17-2, 2-17-1, 1-16-3, 3-16-1, 2-16-2, 1-15-4, 4-15-1, 2-15-3, 3-15-2, 1-14-5, 2-14-4, 4-14-2, 3-14-3, 1-13-6, 6-13-1, 2-13-5, 5-13-2, 3-13-4, 4-13-3, 1-12-7, 7-12-1, 2-12-6, 6-12-2, 3-12-5, 5-12-3, 4-12-4, 1-11-8, 8-11-1, 2-11-7, 7-11-2, 3-11-6, 6-11-3, 4-11-5, 5-11-4, 1-19-1, 1-18-2, 2-18-1, 1-17-3, 3-17-1, 2-17-2, 1-16-4, 4-16-1, 2-16-3, 3-16-2, 1-15-5, 2-15-4, 4-15-2, 3-15-3, 1-14-6, 6-14-1, 2-14-5, 5-14-2, 3-14-4, 4-14-3, 1-13-7, 7-13-1, 2-13-6, 6-13-2, 3-13-5, 5-13-3, 4-13-4, 1-12-8, 8-12-1, 2-12-7, 7-12-2, 3-12-6, 6-12-3, 4-12-5, 5-12-4, 2-11-8, 8-11-2, 3-11-7, 7-11-3, 4-11-6, 6-11-4, 5-11-5, 1-20-1, 1-19-2, 2-19-1, 1-18-3, 3-18-1, 2-18-2, 1-17-4, 4-17-1, 2-17-3, 3-17-2, 1-16-5, 2-16-4, 4-16-2, 3-16-3, 1-15-6, 6-15-1, 2-15-5, 5-15-2, 3-15-4, 4-15-3, 1-14-7, 7-14-1, 2-14-6, 6-14-2, 3-14-5, 5-14-3, 4-14-4, 1-13-8, 8-13-1, 2-13-7, 7-13-2, 3-13-6, 6-13-3, 4-13-5, 5-13-4, 2-12-8, 8-12-2, 3-12-7, 7-12-3, 4-12-6, 6-12-4, 5-12-5, 3-11-8, 8-11-3, 4-11-7, 7-11-4, 5-11-6, 6-11-5, 1-21-1, 1-20-2, 2-20-1, 1-20-3, 3-19-1, 2-19-2, 1-18-4, 4-18-1, 2-18-3, 3-18-2, 1-17-5, 2-17-4, 4-17-2, 3-17-3, 1-16-6, 6-16-1, 2-16-5, 5-16-2, 3-16-4, 4-16-3, 1-15-7, 7-15-1, 2-15-6, 6-15-2, 3-15-5, 5-15-3, 4-15-4, 1-14-8, 8-14-1, 2-14-7, 7-14-2, 3-14-6, 6-14-3, 4-14-5, 5-14-4, 2-13-8, 8-13-2, 3-13-7, 7-13-3, 4-13-6, 6-13-4, 5-13-5, 1-12-10, 10-12-1, 2-12-9, 9-12-2, 3-12-8, 8-12-3, 4-12-7, 7-12-4, 5-12-6, 6-12-5, 4-11-8, 8-11-4, 5-11-7, 7-11-5, 6-11-6, 1-22-1, 1-21-2, 2-21-1, 1-21-3, 3-20-1, 2-20-2, 1-19-4, 4-19-1, 2-19-3, 3-19-2, 1-18-5, 2-18-4, 4-18-2, 3-18-3, 1-17-6, 6-17-1, 2-17-5, 5-17-2, 3-17-4, 4-17-3, 1-16-7, 7-16-1, 2-16-6, 6-16-2, 3-16-5, 5-16-3, 4-16-4, 1-15-8, 8-15-1, 2-15-7, 7-15-2, 3-15-6, 6-15-3, 4-15-5, 5-15-4, 2-14-8, 8-14-2, 3-14-7, 7-14-3, 4-14-6, 6-14-4, 5-14-5, 3-13-8, 8-13-3, 4-13-7, 7-13-4, 5-13-6, 6-13-5, 4-12-8, 8-12-4, 5-12-7, 7-12-5, 6-12-6, 5-11-8, 8-11-5, 6-11-7, or 7-11-6. In certain preferred embodiments, a gap-widened motif includes, but is not limited to, 2-13-5, 3-14-3, 3-14-4 gapmer motif.

In one embodiment, a gap-widened antisense oligonucleotide targeted to a PCSK9 nucleic acid has a gap segment of fourteen 2′-deoxyribonucleotides positioned between wing segments of three chemically modified nucleosides. In one embodiment, the chemical modification comprises a 2′-sugar modification. In another embodiment, the chemical modification comprises a 2′-MOE sugar modification.

In one embodiment, antisense compounds targeted to a PCSK9 nucleic acid possess a 5-10-5 gapmer motif.

Target Nucleic Acids, Target Regions and Nucleotide Sequences

Nucleotide sequences that encode PCSK9 include, without limitation, the following: GENBANK® Accession No. NM_(—)174936.2, first deposited with GENBANK® on Jun. 1, 2003, and incorporated herein as SEQ ID NO: 1; nucleotides 25475000 to 25504000 of GENBANK® Accession No. NT_(—)032977.8, first deposited with GENBANK® on Feb. 26, 2006, and incorporated herein as SEQ ID NO: 2; and GENBANK® Accession No. AK124635.1, first deposited with GENBANK® on Sep. 8, 2003, and incorporated herein as SEQ ID NO: 3.

It is noted that some portions of these nucleotide sequences share identical sequence. For example, portions of SEQ ID NO: 1 are identical to portions of SEQ ID NO: 2; portions of SEQ ID NO: 1 are identical to portions of SEQ ID NO: 3; and portions of SEQ ID NO: 2 are identical to portions of SEQ ID NO: 3. Accordingly, antisense compounds targeted to SEQ ID NO: 1 may also target SEQ ID NO: 2 and/or SEQ ID NO: 3; antisense compounds targeted to SEQ ID NO: 2 may also target SEQ ID NO: 1 and/or SEQ ID NO: 3; and antisense compounds targeted to SEQ ID NO: 3 may also target SEQ ID NO: 1 and/or SEQ ID NO: 2. Examples of such antisense compounds are shown in the following tables.

In certain embodiments, antisense compounds target a PCSK9 nucleic acid having the sequence of GENBANK® Accession No. NM_(—)174936.2, first deposited with GENBANK® on Jun. 1, 2003, and incorporated herein as SEQ ID NO: 1. In certain such embodiments, an antisense oligonucleotide targets SEQ ID NO: 1. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 1 is at least 90% complementary to SEQ ID NO: 1. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 1 is at least 95% complementary to SEQ ID NO: 1. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 1 is 100% complementary to SEQ ID NO: 1. In certain embodiments, an antisense oligonucleotide targeted to SEQ ID NO: 1 comprises a nucleotide sequence selected from the nucleotide sequences set forth in Table 2.

TABLE 2 Nucleotide sequences targeted to NM_174936.2 (SEQ ID NO: 1) 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ ID NO NO: 1 NO: 1 Sequence (5′-3′) 4 135 154 GCGCGGAATCCTGGCTGGGA 5 242 261 GAGGAGACCTAGAGGCCGTG 159 294 313 GCCTGGAGCTGACGGTGCCC 160 298 317 GACCGCCTGGAGCTGACGGT 6 300 319 AGGACCGCCTGGAGCTGACG 162 406 425 AAGGCTAGCACCAGCTCCTC 163 407 426 CAAGGCTAGCACCAGCTCCT 164 408 427 GCAAGGCTAGCACCAGCTCC 165 409 428 CGCAAGGCTAGCACCAGCTC 7 410 429 ACGCAAGGCTAGCACCAGCT 166 411 430 AACGCAAGGCTAGCACCAGC 167 412 431 GAACGCAAGGCTAGCACCAG 168 413 432 GGAACGCAAGGCTAGCACCA 169 414 433 CGGAACGCAAGGCTAGCACC 8 417 436 CCTCGGAACGCAAGGCTAGC 170 421 440 TCCTCCTCGGAACGCAAGGC 171 446 465 GTGCTCGGGTGCTTCGGCCA 172 466 485 TGGAAGGTGGCTGTGGTTCC 9 480 499 CCTTGGCGCAGCGGTGGAAG 173 482 501 ATCCTTGGCGCAGCGGTGGA 174 484 503 GGATCCTTGGCGCAGCGGTG 175 488 507 CCACGGATCCTTGGCGCAGC 176 507 526 CGTAGGTGCCAGGCAACCTC 177 545 564 TGACTGCGAGAGGTGGGTCT 178 555 574 CAGTGCGCTCTGACTGCGAG 179 557 576 GGCAGTGCGCTCTGACTGCG 180 559 578 CGGGCAGTGCGCTCTGACTG 10 561 580 GGCGGGCAGTGCGCTCTGAC 181 562 581 CGGCGGGCAGTGCGCTCTGA 182 591 610 ATCCCCGGCGGGCAGCCTGG 183 595 614 AGGTATCCCCGGCGGGCAGC 184 597 616 TGAGGTATCCCCGGCGGGCA 185 598 617 GTGAGGTATCCCCGGCGGGC 186 599 618 GGTGAGGTATCCCCGGCGGG 11 600 619 TGGTGAGGTATCCCCGGCGG 187 601 620 TTGGTGAGGTATCCCCGGCG 188 602 621 CTTGGTGAGGTATCCCCGGC 189 603 622 TCTTGGTGAGGTATCCCCGG 190 604 623 ATCTTGGTGAGGTATCCCCG 191 605 624 GATCTTGGTGAGGTATCCCC 12 606 625 GGATCTTGGTGAGGTATCCC 192 607 626 AGGATCTTGGTGAGGTATCC 193 609 628 GCAGGATCTTGGTGAGGTAT 194 611 630 ATGCAGGATCTTGGTGAGGT 195 613 632 ACATGCAGGATCTTGGTGAG 13 615 634 AGACATGCAGGATCTTGGTG 196 617 636 GAAGACATGCAGGATCTTGG 14 620 639 ATGGAAGACATGCAGGATCT 197 628 647 AGAAGGCCATGGAAGACATG 198 638 657 GAAGCCAGGAAGAAGGCCAT 15 646 665 TTCACCAGGAAGCCAGGAAG 199 648 667 TCTTCACCAGGAAGCCAGGA 16 651 670 TCATCTTCACCAGGAAGCCA 200 653 672 ACTCATCTTCACCAGGAAGC 201 655 674 CCACTCATCTTCACCAGGAA 202 657 676 CGCCACTCATCTTCACCAGG 203 659 678 GTCGCCACTCATCTTCACCA 204 661 680 AGGTCGCCACTCATCTTCAC 205 663 682 GCAGGTCGCCACTCATCTTC 206 665 684 CAGCAGGTCGCCACTCATCT 207 667 686 TCCAGCAGGTCGCCACTCAT 208 685 704 GGCAACTTCAAGGCCAGCTC 17 705 724 CCTCGATGTAGTCGACATGG 209 724 743 GCAAAGACAGAGGAGTCCTC 210 782 801 TTCATCCGCCCGGTACCGTG 211 784 803 TATTCATCCGCCCGGTACCG 18 785 804 GTATTCATCCGCCCGGTACC 212 787 806 TGGTATTCATCCGCCCGGTA 213 789 808 GCTGGTATTCATCCGCCCGG 214 791 810 GGGCTGGTATTCATCCGCCC 215 821 840 ATACACCTCCACCAGGCTGC 216 832 851 GTGTCTAGGAGATACACCTC 19 835 854 CTGGTGTCTAGGAGATACAC 217 837 856 TGCTGGTGTCTAGGAGATAC 20 840 859 GTATGCTGGTGTCTAGGAGA 21 860 879 GATTTCCCGGTGGTCACTCT 218 862 881 TCGATTTCCCGGTGGTCACT 219 863 882 CTCGATTTCCCGGTGGTCAC 220 864 883 CCTCGATTTCCCGGTGGTCA 221 865 884 CCCTCGATTTCCCGGTGGTC 22 866 885 GCCCTCGATTTCCCGGTGGT 222 867 886 TGCCCTCGATTTCCCGGTGG 223 868 887 CTGCCCTCGATTTCCCGGTG 224 869 888 CCTGCCCTCGATTTCCCGGT 225 870 889 CCCTGCCCTCGATTTCCCGG 226 874 893 ATGACCCTGCCCTCGATTTC 227 876 895 CCATGACCCTGCCCTCGATT 228 878 897 GACCATGACCCTGCCCTCGA 23 880 899 GTGACCATGACCCTGCCCTC 229 882 901 CGGTGACCATGACCCTGCCC 230 884 903 GTCGGTGACCATGACCCTGC 231 886 905 AAGTCGGTGACCATGACCCT 232 888 907 CGAAGTCGGTGACCATGACC 24 890 909 CTCGAAGTCGGTGACCATGA 233 898 917 GGCACATTCTCGAAGTCGGT 25 923 942 GTGGAAGCGGGTCCCGTCCT 234 933 952 TGGCCTGTCTGTGGAAGCGG 235 960 979 GGTGGGTGCCATGACTGTCA 236 963 982 CCAGGTGGGTGCCATGACTG 237 967 986 CCTGCCAGGTGGGTGCCATG 26 970 989 ACCCCTGCCAGGTGGGTGCC 238 972 991 CCACCCCTGCCAGGTGGGTG 27 975 994 TGACCACCCCTGCCAGGTGG 239 977 996 GCTGACCACCCCTGCCAGGT 240 985 1004 TCCCGGCCGCTGACCACCCC 241 989 1008 GGCATCCCGGCCGCTGACCA 242 992 1011 GCCGGCATCCCGGCCGCTGA 243 997 1016 GCCACGCCGGCATCCCGGCC 244 998 1017 GGCCACGCCGGCATCCCGGC 245 999 1018 TGGCCACGCCGGCATCCCGG 246 1000 1019 TTGGCCACGCCGGCATCCCG 247 1001 1020 CTTGGCCACGCCGGCATCCC 248 1002 1021 CCTTGGCCACGCCGGCATCC 249 1003 1022 CCCTTGGCCACGCCGGCATC 28 1004 1023 ACCCTTGGCCACGCCGGCAT 447 1004 1023 ACCCTTGGTCACGCCGGCAT 250 1005 1024 CACCCTTGGCCACGCCGGCA 251 1006 1025 GCACCCTTGGCCACGCCGGC 252 1007 1026 GGCACCCTTGGCCACGCCGG 253 1008 1027 TGGCACCCTTGGCCACGCCG 254 1009 1028 CTGGCACCCTTGGCCACGCC 255 1010 1029 GCTGGCACCCTTGGCCACGC 256 1015 1034 CGCATGCTGGCACCCTTGGC 257 1036 1055 CAGTTGAGCACGCGCAGGCT 258 1038 1057 GGCAGTTGAGCACGCGCAGG 29 1040 1059 TTGGCAGTTGAGCACGCGCA 259 1042 1061 CCTTGGCAGTTGAGCACGCG 30 1045 1064 TTCCCTTGGCAGTTGAGCAC 260 1047 1066 CCTTCCCTTGGCAGTTGAGC 261 1051 1070 GTGCCCTTCCCTTGGCAGTT 262 1053 1072 CCGTGCCCTTCCCTTGGCAG 263 1064 1083 GGTGCCGCTAACCGTGCCCT 264 1076 1095 CAGGCCTATGAGGGTGCCGC 31 1077 1096 CCAGGCCTATGAGGGTGCCG 458 1079 1092 GCCTATGAGGGTGC 459 1084 1097 TCCAGGCCTATGAG 265 1088 1107 CCGAATAAACTCCAGGCCTA 266 1096 1115 TGGCTTTTCCGAATAAACTC 32 1098 1117 GCTGGCTTTTCCGAATAAAC 267 1100 1119 CAGCTGGCTTTTCCGAATAA 268 1102 1121 ACCAGCTGGCTTTTCCGAAT 269 1104 1123 GGACCAGCTGGCTTTTCCGA 270 1108 1127 GGCTGGACCAGCTGGCTTTT 271 1119 1138 GTGGCCCCACAGGCTGGACC 272 1132 1151 AGCAGCACCACCAGTGGCCC 273 1154 1173 GCTGTACCCACCCGCCAGGG 274 1200 1219 CGACCCCAGCCCTCGCCAGG 33 1210 1229 GTGACCAGCACGACCCCAGC 275 1212 1231 CGGTGACCAGCACGACCCCA 276 1214 1233 AGCGGTGACCAGCACGACCC 277 1216 1235 GCAGCGGTGACCAGCACGAC 278 1218 1237 CGGCAGCGGTGACCAGCACG 279 1219 1238 CCGGCAGCGGTGACCAGCAC 280 1222 1241 TTGCCGGCAGCGGTGACCAG 281 1224 1243 AGTTGCCGGCAGCGGTGACC 282 1226 1245 GAAGTTGCCGGCAGCGGTGA 283 1228 1247 CGGAAGTTGCCGGCAGCGGT 284 1230 1249 CCCGGAAGTTGCCGGCAGCG 285 1232 1251 GTCCCGGAAGTTGCCGGCAG 286 1273 1292 ATGACCTCGGGAGCTGAGGC 287 1283 1302 CCCAACTGTGATGACCTCGG 288 1295 1314 GGCATTGGTGGCCCCAACTG 149 1297 1316 TGGGCATTGGTGGCCCCAAC 289 1305 1324 GCTGGTCTTGGGCATTGGTG 290 1318 1337 CCCAGGGTCACCGGCTGGTC 291 1320 1339 TCCCCAGGGTCACCGGCTGG 292 1322 1341 AGTCCCCAGGGTCACCGGCT 293 1324 1343 AAAGTCCCCAGGGTCACCGG 34 1326 1345 CCAAAGTCCCCAGGGTCACC 294 1328 1347 CCCCAAAGTCCCCAGGGTCA 128 1330 1349 GTCCCCAAAGTCCCCAGGGT 295 1333 1352 TTGGTCCCCAAAGTCCCCAG 35 1335 1354 AGTTGGTCCCCAAAGTCCCC 296 1337 1356 AAAGTTGGTCCCCAAAGTCC 36 1340 1359 GCCAAAGTTGGTCCCCAAAG 297 1342 1361 CGGCCAAAGTTGGTCCCCAA 298 1344 1363 AGCGGCCAAAGTTGGTCCCC 299 1346 1365 ACAGCGGCCAAAGTTGGTCC 300 1348 1367 ACACAGCGGCCAAAGTTGGT 301 1350 1369 CCACACAGCGGCCAAAGTTG 37 1352 1371 GTCCACACAGCGGCCAAAGT 302 1354 1373 AGGTCCACACAGCGGCCAAA 303 1356 1375 AGAGGTCCACACAGCGGCCA 304 1358 1377 AAAGAGGTCCACACAGCGGC 38 1361 1380 GGCAAAGAGGTCCACACAGC 305 1380 1399 CAATGATGTCCTCCCCTGGG 306 1387 1406 GAGGCACCAATGATGTCCTC 39 1389 1408 TGGAGGCACCAATGATGTCC 307 1391 1410 GCTGGAGGCACCAATGATGT 308 1393 1412 TCGCTGGAGGCACCAATGAT 309 1395 1414 AGTCGCTGGAGGCACCAATG 310 1397 1416 GCAGTCGCTGGAGGCACCAA 40 1400 1419 GCTGCAGTCGCTGGAGGCAC 311 1402 1421 GTGCTGCAGTCGCTGGAGGC 312 1404 1423 AGGTGCTGCAGTCGCTGGAG 313 1406 1425 GCAGGTGCTGCAGTCGCTGG 314 1407 1426 AGCAGGTGCTGCAGTCGCTG 315 1409 1428 AAAGCAGGTGCTGCAGTCGC 41 1411 1430 ACAAAGCAGGTGCTGCAGTC 316 1413 1432 ACACAAAGCAGGTGCTGCAG 317 1415 1434 TGACACAAAGCAGGTGCTGC 318 1425 1444 TCCCACTCTGTGACACAAAG 101 1465 1484 ATGGCTGCAATGCCAGCCAC 319 1467 1486 TCATGGCTGCAATGCCAGCC 42 1470 1489 GCATCATGGCTGCAATGCCA 320 1472 1491 CAGCATCATGGCTGCAATGC 321 1474 1493 GACAGCATCATGGCTGCAAT 322 1476 1495 CAGACAGCATCATGGCTGCA 43 1478 1497 GGCAGACAGCATCATGGCTG 323 1480 1499 TCGGCAGACAGCATCATGGC 324 1482 1501 GCTCGGCAGACAGCATCATG 325 1484 1503 CGGCTCGGCAGACAGCATCA 326 1486 1505 TCCGGCTCGGCAGACAGCAT 327 1500 1519 CGGCCAGGGTGAGCTCCGGC 328 1513 1532 CTCTGCCTCAACTCGGCCAG 329 1515 1534 GTCTCTGCCTCAACTCGGCC 330 1517 1536 CAGTCTCTGCCTCAACTCGG 331 1519 1538 ATCAGTCTCTGCCTCAACTC 332 1521 1540 GGATCAGTCTCTGCCTCAAC 333 1523 1542 GTGGATCAGTCTCTGCCTCA 334 1525 1544 AAGTGGATCAGTCTCTGCCT 44 1526 1545 GAAGTGGATCAGTCTCTGCC 335 1528 1547 GAGAAGTGGATCAGTCTCTG 336 1530 1549 CAGAGAAGTGGATCAGTCTC 337 1532 1551 GGCAGAGAAGTGGATCAGTC 45 1534 1553 TTGGCAGAGAAGTGGATCAG 338 1536 1555 CTTTGGCAGAGAAGTGGATC 46 1539 1558 CATCTTTGGCAGAGAAGTGG 339 1541 1560 GACATCTTTGGCAGAGAAGT 340 1543 1562 ATGACATCTTTGGCAGAGAA 47 1545 1564 TGATGACATCTTTGGCAGAG 341 1547 1566 ATTGATGACATCTTTGGCAG 342 1549 1568 TCATTGATGACATCTTTGGC 48 1552 1571 GCCTCATTGATGACATCTTT 343 1554 1573 AGGCCTCATTGATGACATCT 344 1556 1575 CCAGGCCTCATTGATGACAT 345 1558 1577 AACCAGGCCTCATTGATGAC 346 1560 1579 GGAACCAGGCCTCATTGATG 347 1561 1580 GGGAACCAGGCCTCATTGAT 348 1562 1581 AGGGAACCAGGCCTCATTGA 349 1563 1582 CAGGGAACCAGGCCTCATTG 49 1564 1583 TCAGGGAACCAGGCCTCATT 49 1564 1583 TCAGGGAACCAGGCCTCATT 350 1565 1584 CTCAGGGAACCAGGCCTCAT 351 1566 1585 CCTCAGGGAACCAGGCCTCA 352 1567 1586 TCCTCAGGGAACCAGGCCTC 353 1568 1587 GTCCTCAGGGAACCAGGCCT 50 1569 1588 GGTCCTCAGGGAACCAGGCC 354 1570 1589 TGGTCCTCAGGGAACCAGGC 355 1571 1590 CTGGTCCTCAGGGAACCAGG 356 1572 1591 GCTGGTCCTCAGGGAACCAG 357 1573 1592 CGCTGGTCCTCAGGGAACCA 87 1576 1595 ACCCGCTGGTCCTCAGGGAA 358 1578 1597 GTACCCGCTGGTCCTCAGGG 359 1580 1599 CAGTACCCGCTGGTCCTCAG 51 1583 1602 GGTCAGTACCCGCTGGTCCT 119 1605 1624 GCAGGGCGGCCACCAGGTTG 360 1628 1647 ACCTGCCCCATGGGTGCTGG 52 1640 1659 AAACAGCTGCCAACCTGCCC 361 1642 1661 CAAAACAGCTGCCAACCTGC 53 1645 1664 CTGCAAAACAGCTGCCAACC 362 1647 1666 TCCTGCAAAACAGCTGCCAA 363 1649 1668 AGTCCTGCAAAACAGCTGCC 364 1660 1679 GCTGACCATACAGTCCTGCA 365 1672 1691 GGCCCCGAGTGTGCTGACCA 54 1675 1694 GTAGGCCCCGAGTGTGCTGA 366 1677 1696 GTGTAGGCCCCGAGTGTGCT 367 1679 1698 CCGTGTAGGCCCCGAGTGTG 368 1681 1700 ATCCGTGTAGGCCCCGAGTG 369 1683 1702 CCATCCGTGTAGGCCCCGAG 370 1685 1704 GGCCATCCGTGTAGGCCCCG 371 1687 1706 GTGGCCATCCGTGTAGGCCC 372 1735 1754 CTGGAGCAGCTCAGCAGCTC 460 1735 1748 CAGCTCAGCAGCTC 373 1737 1756 AACTGGAGCAGCTCAGCAGC 55 1740 1759 AGAAACTGGAGCAGCTCAGC 374 1742 1761 GGAGAAACTGGAGCAGCTCA 375 1744 1763 CTGGAGAAACTGGAGCAGCT 56 1746 1765 TCCTGGAGAAACTGGAGCAG 57 1812 1831 CGTTGTGGGCCCGGCAGACC 376 1849 1868 CACCTGGCAATGGCGTAGAC 377 1850 1869 GCACCTGGCAATGGCGTAGA 378 1851 1870 AGCACCTGGCAATGGCGTAG 379 1852 1871 CAGCACCTGGCAATGGCGTA 380 1853 1872 GCAGCACCTGGCAATGGCGT 381 1854 1873 GGCAGCACCTGGCAATGGCG 382 1855 1874 AGGCAGCACCTGGCAATGGC 383 1856 1875 CAGGCAGCACCTGGCAATGG 384 1857 1876 GCAGGCAGCACCTGGCAATG 58 1858 1877 AGCAGGCAGCACCTGGCAAT 385 1859 1878 TAGCAGGCAGCACCTGGCAA 386 1860 1879 GTAGCAGGCAGCACCTGGCA 387 1905 1924 TGGCCTCAGCTGGTGGAGCT 388 1915 1934 GTCCCCATGCTGGCCTCAGC 389 1916 1935 GGTCCCCATGCTGGCCTCAG 390 1917 1936 GGGTCCCCATGCTGGCCTCA 391 1918 1937 CGGGTCCCCATGCTGGCCTC 392 1919 1938 ACGGGTCCCCATGCTGGCCT 59 1920 1939 CACGGGTCCCCATGCTGGCC 59 1920 1939 CACGGGTCCCCATGCTGGCC 393 1921 1940 ACACGGGTCCCCATGCTGGC 394 1922 1941 GACACGGGTCCCCATGCTGG 395 1923 1942 GGACACGGGTCCCCATGCTG 396 1924 1943 TGGACACGGGTCCCCATGCT 397 1925 1944 GTGGACACGGGTCCCCATGC 398 1926 1945 AGTGGACACGGGTCCCCATG 399 1927 1946 CAGTGGACACGGGTCCCCAT 400 1928 1947 GCAGTGGACACGGGTCCCCA 401 1929 1948 GGCAGTGGACACGGGTCCCC 402 1930 1949 TGGCAGTGGACACGGGTCCC 403 1931 1950 GTGGCAGTGGACACGGGTCC 404 1932 1951 GGTGGCAGTGGACACGGGTC 405 1933 1952 TGGTGGCAGTGGACACGGGT 406 1936 1955 TGTTGGTGGCAGTGGACACG 407 1962 1981 AGCTGCAGCCTGTGAGGACG 408 1990 2009 GTGCCAAGGTCCTCCACCTC 409 2010 2029 TCAGCACAGGCGGCTTGTGG 410 2040 2059 CCACGCACTGGTTGGGCTGA 60 2100 2119 CTTTGCATTCCAGACCTGGG 411 2101 2120 ACTTTGCATTCCAGACCTGG 412 2102 2121 GACTTTGCATTCCAGACCTG 413 2103 2122 TGACTTTGCATTCCAGACCT 414 2104 2123 TTGACTTTGCATTCCAGACC 61 2105 2124 CTTGACTTTGCATTCCAGAC 415 2107 2126 TCCTTGACTTTGCATTCCAG 416 2120 2139 CGGGATTCCATGCTCCTTGA 417 2150 2169 GCAGGCCACGGTCACCTGCT 418 2187 2206 GGAGGGCACTGCAGCCAGTC 419 2305 2324 CAGATGGCAACGGCTGTCAC 420 2306 2325 GCAGATGGCAACGGCTGTCA 421 2307 2326 AGCAGATGGCAACGGCTGTC 422 2308 2327 CAGCAGATGGCAACGGCTGT 423 2309 2328 GCAGCAGATGGCAACGGCTG 62 2310 2329 GGCAGCAGATGGCAACGGCT 424 2311 2330 CGGCAGCAGATGGCAACGGC 425 2312 2331 CCGGCAGCAGATGGCAACGG 426 2313 2332 TCCGGCAGCAGATGGCAACG 427 2314 2333 CTCCGGCAGCAGATGGCAAC 428 2315 2334 GCTCCGGCAGCAGATGGCAA 429 2325 2344 CCAGGTGCCGGCTCCGGCAG 430 2335 2354 GAGGCCTGCGCCAGGTGCCG 154 2410 2429 TTTTAAAGCTCAGCCCCAGC 63 2415 2434 AACCATTTTAAAGCTCAGCC 64 2504 2523 TCAAGGGCCAGGCCAGCAGC 65 2509 2528 CCCACTCAAGGGCCAGGCCA 122 2582 2601 GGAGGGAGCTTCCTGGCACC 66 2597 2616 ATGCCCCACAGTGAGGGAGG 67 2606 2625 AATGGTGAAATGCCCCACAG 153 2649 2668 TTGGGAGCAGCTGGCAGCAC 68 2750 2769 CATGGGAAGAATCCTGCCTC 431 2828 2847 ATGAGGGCCATCAGCACCTT 432 2829 2848 GATGAGGGCCATCAGCACCT 433 2830 2849 AGATGAGGGCCATCAGCACC 434 2831 2850 GAGATGAGGGCCATCAGCAC 69 2832 2851 GGAGATGAGGGCCATCAGCA 435 2833 2852 TGGAGATGAGGGCCATCAGC 436 2834 2853 CTGGAGATGAGGGCCATCAG 437 2835 2854 GCTGGAGATGAGGGCCATCA 438 2836 2855 AGCTGGAGATGAGGGCCATC 461 2877 2890 TTAATCAGGGAGCC 70 2900 2919 TAGATGCCATCCAGAAAGCT 439 2902 2921 GCTAGATGCCATCCAGAAAG 440 2903 2922 GGCTAGATGCCATCCAGAAA 441 2904 2923 TGGCTAGATGCCATCCAGAA 442 2905 2924 CTGGCTAGATGCCATCCAGA 71 2906 2925 TCTGGCTAGATGCCATCCAG 443 2907 2926 CTCTGGCTAGATGCCATCCA 444 2908 2927 CCTCTGGCTAGATGCCATCC 445 2909 2928 GCCTCTGGCTAGATGCCATC 446 2910 2929 AGCCTCTGGCTAGATGCCAT 72 2983 3002 GGCATAGAGCAGAGTAAAGG 73 2988 3007 AGCCTGGCATAGAGCAGAGT 135 2994 3013 TAGCACAGCCTGGCATAGAG 112 3227 3246 GAAGAGGCTTGGCTTCAGAG 74 3233 3252 AAGTAAGAAGAGGCTTGGCT 75 3437 3456 GCTCAAGGAGGGACAGTTGT 76 3472 3491 AAAGATAAATGTCTGCTTGC 77 3477 3496 ACCCAAAAGATAAATGTCTG 78 3543 3562 TCTTCAAGTTACAAAAGCAA 99 3550 3569 ATAAATATCTTCAAGTTACA

In certain embodiments, gapmer antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gapmer antisense compounds are targeted to SEQ ID NO: 1. In certain such embodiments, the nucleotide sequences illustrated in Table 2 have a 5-10-5 gapmer motif. Table 3 illustrates gapmer antisense compounds targeted to SEQ ID NO: 1, having a 5-10-5 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 3 Gapmer antisense compounds having a 5-10-5 motif targeted to SEQ ID NO: 1 5′ Target 3′ Target SEQ Site on Site on ID SEQ ID SEQ ID Isis No Motif NO NO: 1 NO: 1 Mismatches Sequence (5′-3′) 395149 5-10-5 4 135 154 0 GCGCGGAATCCTGGCTGGGA 395150 5-10-5 5 242 261 0 GAGGAGACCTAGAGGCCGTG 395151 5-10-5 6 300 319 0 AGGACCGCCTGGAGCTGACG 395152 5-10-5 7 410 429 0 ACGCAAGGCTAGCACCAGCT 399793 5-10-5 8 417 436 0 CCTCGGAACGCAAGGCTAGC 395153 5-10-5 9 480 499 0 CCTTGGCGCAGCGGTGGAAG 395154 5-10-5 10 561 580 0 GGCGGGCAGTGCGCTCTGAC 395155 5-10-5 11 600 619 0 TGGTGAGGTATCCCCGGCGG 399794 5-10-5 12 606 625 0 GGATCTTGGTGAGGTATCCC 399795 5-10-5 13 615 634 0 AGACATGCAGGATCTTGGTG 395156 5-10-5 14 620 639 0 ATGGAAGACATGCAGGATCT 395157 5-10-5 15 646 665 0 TTCACCAGGAAGCCAGGAAG 399796 5-10-5 16 651 670 0 TCATCTTCACCAGGAAGCCA 395158 5-10-5 17 705 724 0 CCTCGATGTAGTCGACATGG 395159 5-10-5 18 785 804 0 GTATTCATCCGCCCGGTACC 395160 5-10-5 19 835 854 0 CTGGTGTCTAGGAGATACAC 399797 5-10-5 20 840 859 0 GTATGCTGGTGTCTAGGAGA 395161 5-10-5 21 860 879 0 GATTTCCCGGTGGTCACTCT 399798 5-10-5 22 866 885 0 GCCCTCGATTTCCCGGTGGT 399799 5-10-5 23 880 899 0 GTGACCATGACCCTGCCCTC 395162 5-10-5 24 890 909 0 CTCGAAGTCGGTGACCATGA 395163 5-10-5 25 923 942 0 GTGGAAGCGGGTCCCGTCCT 395164 5-10-5 26 970 989 0 ACCCCTGCCAGGTGGGTGCC 399800 5-10-5 27 975 994 0 TGACCACCCCTGCCAGGTGG 395165 5-10-5 28 1004 1023 0 ACCCTTGGCCACGCCGGCAT 395166 5-10-5 29 1040 1059 0 TTGGCAGTTGAGCACGCGCA 399801 5-10-5 30 1045 1064 0 TTCCCTTGGCAGTTGAGCAC 395167 5-10-5 31 1077 1096 0 CCAGGCCTATGAGGGTGCCG 395168 5-10-5 32 1098 1117 0 GCTGGCTTTTCCGAATAAAC 395169 5-10-5 33 1210 1229 0 GTGACCAGCACGACCCCAGC 395170 5-10-5 149 1297 1316 0 TGGGCATTGGTGGCCCCAAC 395171 5-10-5 34 1326 1345 0 CCAAAGTCCCCAGGGTCACC 395172 5-10-5 128 1330 1349 0 GTCCCCAAAGTCCCCAGGGT 399802 5-10-5 35 1335 1354 0 AGTTGGTCCCCAAAGTCCCC 395173 5-10-5 36 1340 1359 0 GCCAAAGTTGGTCCCCAAAG 399803 5-10-5 37 1352 1371 0 GTCCACACAGCGGCCAAAGT 395174 5-10-5 38 1361 1380 0 GGCAAAGAGGTCCACACAGC 395175 5-10-5 39 1389 1408 0 TGGAGGCACCAATGATGTCC 399804 5-10-5 40 1400 1419 0 GCTGCAGTCGCTGGAGGCAC 399805 5-10-5 41 1411 1430 0 ACAAAGCAGGTGCTGCAGTC 395176 5-10-5 101 1465 1484 0 ATGGCTGCAATGCCAGCCAC 399806 5-10-5 42 1470 1489 0 GCATCATGGCTGCAATGCCA 399807 5-10-5 43 1478 1497 0 GGCAGACAGCATCATGGCTG 399808 5-10-5 44 1526 1545 0 GAAGTGGATCAGTCTCTGCC 395177 5-10-5 45 1534 1553 0 TTGGCAGAGAAGTGGATCAG 399809 5-10-5 46 1539 1558 0 CATCTTTGGCAGAGAAGTGG 399810 5-10-5 47 1545 1564 0 TGATGACATCTTTGGCAGAG 399811 5-10-5 48 1552 1571 0 GCCTCATTGATGACATCTTT 399812 5-10-5 49 1564 1583 0 TCAGGGAACCAGGCCTCATT 395178 5-10-5 50 1569 1588 0 GGTCCTCAGGGAACCAGGCC 395179 5-10-5 87 1576 1595 0 ACCCGCTGGTCCTCAGGGAA 399813 5-10-5 51 1583 1602 0 GGTCAGTACGCGCTGGTCCT 395180 5-10-5 119 1605 1624 0 GCAGGGCGGCCACCAGGTTG 395181 5-10-5 52 1640 1659 0 AAACAGCTGCCAACCTGCCC 399814 5-10-5 53 1645 1664 0 CTGCAAAACAGCTGCCAACC 395182 5-10-5 54 1675 1694 0 GTAGGCCCCGAGTGTGCTGA 399815 5-10-5 55 1740 1759 0 AGAAACTGGAGCAGCTCAGC 399816 5-10-5 56 1746 1765 0 TCCTGGAGAAACTGGAGCAG 395183 5-10-5 57 1812 1831 0 CGTTGTGGGCCCGGCAGACC 395184 5-10-5 58 1858 1877 0 AGCAGGCAGCACCTGGCAAT 395185 5-10-5 59 1920 1939 0 CACGGGTCCCCATGCTGGCC 395186 5-10-5 60 2100 2119 0 CTTTGCATTCCAGACCTGGG 399817 5-10-5 61 2105 2124 0 CTTGACTTTGCATTCCAGAC 395187 5-10-5 62 2310 2329 0 GGCAGCAGATGGCAACGGCT 395188 5-10-5 154 2410 2429 0 TTTTAAAGCTCAGCCCCAGC 399818 5-10-5 63 2415 2434 0 AACCATTTTAAAGCTCAGCC 395189 5-10-5 64 2504 2523 0 TCAAGGGCCAGGCCAGCAGC 399819 5-10-5 65 2509 2528 0 CCCACTCAAGGGCCAGGCCA 399820 5-10-5 122 2582 2601 0 GGAGGGAGCTTCCTGGCACC 395190 5-10-5 66 2597 2616 0 ATGCCCCACAGTGAGGGAGG 395191 5-10-5 67 2606 2625 0 AATGGTGAAATGCCCCACAG 395192 5-10-5 153 2649 2668 0 TTGGGAGCAGCTGGCAGCAC 395193 5-10-5 68 2750 2769 0 CATGGGAAGAATCCTGCCTC 395194 5-10-5 69 2832 2851 0 GGAGATGAGGGCCATCAGCA 395195 5-10-5 70 2900 2919 0 TAGATGCCATCCAGAAAGCT 399821 5-10-5 71 2906 2925 0 TCTGGCTAGATGCCATCCAG 395196 5-10-5 72 2983 3002 0 GGCATAGAGCAGAGTAAAGG 399822 5-10-5 73 2988 3007 0 AGCCTGGCATAGAGCAGAGT 399823 5-10-5 135 2994 3013 0 TAGCACAGCCTGGCATAGAG 395197 5-10-5 112 3227 3246 0 GAAGAGGCTTGGCTTCAGAG 399824 5-10-5 74 3233 3252 0 AAGTAAGAAGAGGCTTGGCT 395198 5-10-5 75 3437 3456 0 GCTCAAGGAGGGACAGTTGT 395199 5-10-5 76 3472 3491 0 AAAGATAAATGTCTGCTTGC 399825 5-10-5 77 3477 3496 0 ACCCAAAAGATAAATGTCTG 395200 5-10-5 78 3543 3562 0 TCTTCAAGTTACAAAAGCAA 399826 5-10-5 99 3550 3569 0 ATAAATATCTTCAAGTTACA 405861 5-10-5 162 406 425 0 AAGGCTAGCACCAGCTCCTC 405862 5-10-5 163 407 426 0 CAAGGCTAGCACCAGCTCCT 405863 5-10-5 164 408 427 0 GCAAGGCTAGCACCAGCTCC 405864 5-10-5 165 409 428 0 CGCAAGGCTAGCACCAGCTC 405865 5-10-5 166 411 430 0 AACGCAAGGCTAGCACCAGC 405866 5-10-5 167 412 431 0 GAACGCAAGGCTAGCACCAG 405867 5-10-5 168 413 432 0 GGAACGCAAGGCTAGCACCA 405868 5-10-5 169 414 433 0 CGGAACGCAAGGCTAGCACC 405869 5-10-5 218 862 881 0 TCGATTTCCCGGTGGTCACT 405870 5-10-5 219 863 882 0 CTCGATTTCCCGGTGGTCAC 405871 5-10-5 220 864 883 0 CCTCGATTTCCCGGTGGTCA 405872 5-10-5 221 865 884 0 CCCTCGATTTCCCGGTGGTC 405873 5-10-5 222 867 886 0 TGCCCTCGATTTCCCGGTGG 405874 5-10-5 223 868 887 0 CTGCCCTCGATTTCCCGGTG 405875 5-10-5 224 869 888 0 CCTGCCCTCGATTTCCCGGT 405876 5-10-5 225 870 889 0 CCCTGCCCTCGATTTCCCGG 405877 5-10-5 246 1000 1019 0 TTGGCCACGCCGGCATCCCG 405878 5-10-5 247 1001 1020 0 CTTGGCCACGCCGGCATCCC 405879 5-10-5 248 1002 1021 0 CCTTGGCCACGCCGGCATCC 405880 5-10-5 249 1003 1022 0 CCCTTGGCCACGCCGGCATC 405881 5-10-5 250 1005 1024 0 CACCCTTGGCCACGCCGGCA 405882 5-10-5 251 1006 1025 0 GCACCCTTGGCCACGCCGGC 405883 5-10-5 252 1007 1026 0 GGCACCCTTGGCCACGCCGG 405884 5-10-5 253 1008 1027 0 TGGCACCCTTGGCCACGCCG 405885 5-10-5 346 1560 1579 0 GGAACCAGGCCTCATTGATG 405886 5-10-5 347 1561 1580 0 GGGAACCAGGCCTCATTGAT 405887 5-10-5 348 1562 1581 0 AGGGAACCAGGCCTCATTGA 405888 5-10-5 349 1563 1582 0 CAGGGAACCAGGCCTCATTG 405889 5-10-5 350 1565 1584 0 CTCAGGGAACCAGGCCTCAT 405890 5-10-5 351 1566 1585 0 CCTCAGGGAACCAGGCCTCA 405891 5-10-5 352 1567 1586 0 TCCTCAGGGAACCAGGCCTC 405892 5-10-5 353 1568 1587 0 GTCCTCAGGGAACCAGGCCT 405893 5-10-5 431 2828 2847 0 ATGAGGGCCATCAGCACCTT 405894 5-10-5 432 2829 2848 0 GATGAGGGCCATCAGCACCT 405895 5-10-5 433 2830 2849 0 AGATGAGGGCCATCAGCACC 405896 5-10-5 434 2831 2850 0 GAGATGAGGGCCATCAGCAC 405897 5-10-5 435 2833 2852 0 TGGAGATGAGGGCCATCAGC 405898 5-10-5 436 2834 2853 0 CTGGAGATGAGGGCCATCAG 405899 5-10-5 437 2835 2854 0 GCTGGAGATGAGGGCCATCA 405900 5-10-5 438 2836 2855 0 AGCTGGAGATGAGGGCCATC 405901 5-10-5 439 2902 2921 0 GCTAGATGCCATCCAGAAAG 405902 5-10-5 440 2903 2922 0 GGCTAGATGCCATCCAGAAA 405903 5-10-5 441 2904 2923 0 TGGCTAGATGCCATCCAGAA 405904 5-10-5 442 2905 2924 0 CTGGCTAGATGCCATCCAGA 405905 5-10-5 443 2907 2926 0 CTCTGGCTAGATGCCATCCA 405906 5-10-5 444 2908 2927 0 CCTCTGGCTAGATGCCATCC 405907 5-10-5 445 2909 2928 0 GCCTCTGGCTAGATGCCATC 405908 5-10-5 446 2910 2929 0 AGCCTCTGGCTAGATGCCAT 405909 5-10-5 267 1100 1119 0 CAGCTGGCTTTTCCGAATAA 405910 5-10-5 268 1102 1121 0 ACCAGCTGGCTTTTCCGAAT 405911 5-10-5 269 1104 1123 0 GGACCAGCTGGCTTTTCCGA 405912 5-10-5 270 1108 1127 0 GGCTGGACCAGCTGGCTTTT 405913 5-10-5 275 1212 1231 0 CGGTGACCAGCACGACCCCA 405914 5-10-5 276 1214 1233 0 AGCGGTGACCAGCACGACCC 405915 5-10-5 277 1216 1235 0 GCAGCGGTGACCAGCACGAC 405916 5-10-5 278 1218 1237 0 CGGCAGCGGTGACCAGCACG 405917 5-10-5 280 1222 1241 0 TTGCCGGCAGCGGTGACCAG 405918 5-10-5 281 1224 1243 0 AGTTGCCGGCAGCGGTGACC 405919 5-10-5 282 1226 1245 0 GAAGTTGCCGGCAGCGGTGA 405920 5-10-5 283 1228 1247 0 CGGAAGTTGCCGGCAGCGGT 405921 5-10-5 284 1230 1249 0 CCCGGAAGTTGCCGGCAGCG 405922 5-10-5 285 1232 1251 0 GTCCCGGAAGTTGCCGGCAG 405923 5-10-5 288 1295 1314 0 GGCATTGGTGGCCCCAACTG 405924 5-10-5 290 1318 1337 0 CCCAGGGTCACCGGCTGGTC 405925 5-10-5 292 1322 1341 0 AGTCCCCAGGGTCACCGGCT 405926 5-10-5 293 1324 1343 0 AAAGTCCCCAGGGTCACCGG 405927 5-10-5 294 1328 1347 0 CCCCAAAGTCCCCAGGGTCA 405928 5-10-5 295 1333 1352 0 TTGGTCCCCAAAGTCCCCAG 405929 5-10-5 296 1337 1356 0 AAAGTTGGTCCCCAAAGTCC 405930 5-10-5 297 1342 1361 0 CGGCCAAAGTTGGTCCCCAA 405931 5-10-5 298 1344 1363 0 AGCGGCCAAAGTTGGTCCCC 405932 5-10-5 299 1346 1365 0 ACAGCGGCCAAAGTTGGTCC 405933 5-10-5 300 1348 1367 0 ACACAGCGGCCAAAGTTGGT 405934 5-10-5 301 1350 1369 0 CCACACAGCGGCCAAAGTTG 405935 5-10-5 302 1354 1373 0 AGGTCCACACAGCGGCCAAA 405936 5-10-5 304 1358 1377 0 AAAGAGGTCCACACAGCGGC 405937 5-10-5 306 1387 1406 0 GAGGCACCAATGATGTCCTC 405938 5-10-5 307 1391 1410 0 GCTGGAGGCACCAATGATGT 405939 5-10-5 308 1393 1412 0 TCGCTGGAGGCACCAATGAT 405940 5-10-5 309 1395 1414 0 AGTCGCTGGAGGCACCAATG 405941 5-10-5 310 1397 1416 0 GCAGTCGCTGGAGGCACCAA 405942 5-10-5 311 1402 1421 0 GTGCTGCAGTCGCTGGAGGC 405943 5-10-5 312 1404 1423 0 AGGTGCTGCAGTCGCTGGAG 405944 5-10-5 314 1407 1426 0 AGCAGGTGCTGCAGTCGCTG 405945 5-10-5 315 1409 1428 0 AAAGCAGGTGCTGCAGTCGC 405946 5-10-5 316 1413 1432 0 ACACAAAGCAGGTGCTGCAG 405947 5-10-5 317 1415 1434 0 TGACACAAAGCAGGTGCTGC 405948 5-10-5 319 1467 1486 0 TCATGGCTGCAATGCCAGCC 405949 5-10-5 320 1472 1491 0 CAGCATCATGGCTGCAATGC 405950 5-10-5 321 1474 1493 0 GACAGCATCATGGCTGCAAT 405951 5-10-5 322 1476 1495 0 CAGACAGCATCATGGCTGCA 405952 5-10-5 323 1480 1499 0 TCGGCAGACAGCATCATGGC 405953 5-10-5 325 1484 1503 0 CGGCTCGGCAGACAGCATCA 405954 5-10-5 326 1486 1505 0 TCCGGCTCGGCAGACAGCAT 405955 5-10-5 328 1513 1532 0 CTCTGCCTCAACTCGGCCAG 405956 5-10-5 329 1515 1534 0 GTCTCTGCCTCAACTCGGCC 405957 5-10-5 330 1517 1536 0 CAGTCTCTGCCTCAACTCGG 405958 5-10-5 331 1519 1538 0 ATCAGTCTCTGCCTCAACTC 405959 5-10-5 332 1521 1540 0 GGATCAGTCTCTGCCTCAAC 405960 5-10-5 333 1523 1542 0 GTGGATCAGTCTCTGCCTCA 405961 5-10-5 334 1525 1544 0 AAGTGGATCAGTCTCTGCCT 405962 5-10-5 335 1528 1547 0 GAGAAGTGGATCAGTCTCTG 405963 5-10-5 337 1532 1551 0 GGCAGAGAAGTGGATCAGTC 405964 5-10-5 338 1536 1555 0 CTTTGGCAGAGAAGTGGATC 405965 5-10-5 339 1541 1560 0 GACATCTTTGGCAGAGAAGT 405966 5-10-5 340 1543 1562 0 ATGACATCTTTGGCAGAGAA 405967 5-10-5 341 1547 1566 0 ATTGATGACATCTTTGGCAG 405968 5-10-5 342 1549 1568 0 TCATTGATGACATCTTTGGC 405969 5-10-5 343 1554 1573 0 AGGCCTCATTGATGACATCT 405970 5-10-5 344 1556 1575 0 CCAGGCCTCATTGATGACAT 405971 5-10-5 355 1571 1590 0 CTGGTCCTCAGGGAACCAGG 405972 5-10-5 357 1573 1592 0 CGCTGGTCCTCAGGGAACCA 405973 5-10-5 358 1578 1597 0 GTACCCGCTGGTCCTCAGGG 405974 5-10-5 359 1580 1599 0 CAGTACCCGCTGGTCCTCAG 405975 5-10-5 361 1642 1661 0 CAAAACAGCTGCCAACCTGC 405976 5-10-5 362 1647 1666 0 TCCTGCAAAACAGCTGCCAA 405977 5-10-5 363 1649 1668 0 AGTCCTGCAAAACAGCTGCC 405978 5-10-5 365 1672 1691 0 GGCCCCGAGTGTGCTGACCA 405979 5-10-5 366 1677 1696 0 GTGTAGGCCCCGAGTGTGCT 405980 5-10-5 367 1679 1698 0 CCGTGTAGGCCCCGAGTGTG 405981 5-10-5 368 1681 1700 0 ATCCGTGTAGGCCCCGAGTG 405982 5-10-5 370 1685 1704 0 GGCCATCCGTGTAGGCCCCG 405983 5-10-5 371 1687 1706 0 GTGGCCATCCGTGTAGGCCC 405984 5-10-5 372 1735 1754 0 CTGGAGCAGCTCAGCAGCTC 405985 5-10-5 373 1737 1756 0 AACTGGAGCAGCTCAGCAGC 405986 5-10-5 374 1742 1761 0 GGAGAAACTGGAGCAGCTCA 405987 5-10-5 375 1744 1763 0 CTGGAGAAACTGGAGCAGCT 405988 5-10-5 381 1854 1873 0 GGCAGCACCTGGCAATGGCG 405989 5-10-5 383 1856 1875 0 CAGGCAGCACCTGGCAATGG 405990 5-10-5 386 1860 1879 0 GTAGCAGGCAGCACCTGGCA 405991 5-10-5 394 1922 1941 0 GACACGGGTCCCCATGCTGG 405992 5-10-5 396 1924 1943 0 TGGACACGGGTCCCCATGCT 405993 5-10-5 398 1926 1945 0 AGTGGACACGGGTCCCCATG 405994 5-10-5 400 1928 1947 0 GCAGTGGACACGGGTCCCCA 405995 5-10-5 402 1930 1949 0 TGGCAGTGGACACGGGTCCC 405996 5-10-5 412 2102 2121 0 GACTTTGCATTCCAGACCTG 405997 5-10-5 415 2107 2126 0 TCCTTGACTTTGCATTCCAG 405998 5-10-5 426 2313 2332 0 TCCGGCAGCAGATGGCAACG 405999 5-10-5 160 298 317 0 GACCGCCTGGAGCTGACGGT 406000 5-10-5 173 482 501 0 ATCCTTGGCGCAGCGGTGGA 406001 5-10-5 174 484 503 0 GGATCCTTGGCGCAGCGGTG 406002 5-10-5 175 488 507 0 CCACGGATCCTTGGCGCAGC 406003 5-10-5 178 555 574 0 CAGTGCGCTCTGACTGCGAG 406004 5-10-5 179 557 576 0 GGCAGTGCGCTCTGACTGCG 406005 5-10-5 180 559 578 0 CGGGCAGTGCGCTCTGACTG 406006 5-10-5 181 562 581 0 CGGCGGGCAGTGCGCTCTGA 406007 5-10-5 183 595 614 0 AGGTATCCCCGGCGGGCAGC 406008 5-10-5 188 602 621 0 CTTGGTGAGGTATCCCCGGC 406009 5-10-5 190 604 623 0 ATCTTGGTGAGGTATCCCCG 406010 5-10-5 193 609 628 0 GCAGGATCTTGGTGAGGTAT 406011 5-10-5 194 611 630 0 ATGCAGGATCTTGGTGAGGT 406012 5-10-5 195 613 632 0 ACATGCAGGATCTTGGTGAG 406013 5-10-5 196 617 636 0 GAAGACATGCAGGATCTTGG 406014 5-10-5 199 648 667 0 TCTTCACCAGGAAGCCAGGA 406015 5-10-5 200 653 672 0 ACTCATCTTCACCAGGAAGC 406016 5-10-5 201 655 674 0 CCACTCATCTTCACCAGGAA 406017 5-10-5 203 659 678 0 GTCGCCACTCATCTTCACCA 406018 5-10-5 204 661 680 0 AGGTCGCCACTCATCTTCAC 406019 5-10-5 205 663 682 0 GCAGGTCGCCACTCATCTTC 406020 5-10-5 206 665 684 0 CAGCAGGTCGCCACTCATCT 406021 5-10-5 210 782 801 0 TTCATCCGCCCGGTACCGTG 406022 5-10-5 211 784 803 0 TATTCATCCGCCCGGTACCG 406023 5-10-5 212 787 806 0 TGGTATTCATCCGCCCGGTA 406024 5-10-5 213 789 808 0 GCTGGTATTCATCCGCCCGG 406025 5-10-5 216 832 851 0 GTGTCTAGGAGATACACCTC 406026 5-10-5 217 837 856 0 TGCTGGTGTCTAGGAGATAC 406027 5-10-5 226 874 893 0 ATGACCCTGCCCTCGATTTC 406028 5-10-5 227 876 895 0 CCATGACCCTGCCCTCGATT 406029 5-10-5 228 878 897 0 GACCATGACCCTGCCCTCGA 406030 5-10-5 229 882 901 0 CGGTGACCATGACCCTGCCC 406031 5-10-5 230 884 903 0 GTCGGTGACCATGACCCTGC 406032 5-10-5 232 888 907 0 CGAAGTCGGTGACCATGACC 406033 5-10-5 237 967 986 0 CCTGCCAGGTGGGTGCCATG 406034 5-10-5 238 972 991 0 CCACCCCTGCCAGGTGGGTG 406035 5-10-5 239 977 996 0 GCTGACCACCCCTGCCAGGT 406036 5-10-5 241 989 1008 0 GGCATCCCGGCCGCTGACCA 406037 5-10-5 242 992 1011 0 GCCGGCATCCCGGCCGCTGA 406038 5-10-5 245 999 1018 0 TGGCCACGCCGGCATCCCGG 406039 5-10-5 257 1036 1055 0 CAGTTGAGCACGCGCAGGCT 406040 5-10-5 258 1038 1057 0 GGCAGTTGAGCACGCGCAGG 406041 5-10-5 259 1042 1061 0 CCTTGGCAGTTGAGCACGCG 406042 5-10-5 260 1047 1066 0 CCTTCCCTTGGCAGTTGAGC 406043 5-10-5 261 1051 1070 0 GTGCCCTTCCCTTGGCAGTT 406044 5-10-5 262 1053 1072 0 CCGTGCCCTTCCCTTGGCAG 406045 5-10-5 266 1096 1115 0 TGGCTTTTCCGAATAAACTC 408642 5-10-5 264 1076 1095 0 CAGGCCTATGAGGGTGCCGC 408653 5-10-5 354 1570 1589 0 TGGTCCTCAGGGAACCAGGC 409126 5-10-5 447 1004 1023 1 ACCCTTGGTCACGCCGGCAT 410529 5-10-5 184 597 616 0 TGAGGTATCCCCGGCGGGCA 410530 5-10-5 185 598 617 0 GTGAGGTATCCCCGGCGGGC 410531 5-10-5 186 599 618 0 GGTGAGGTATCCCCGGCGGG 410532 5-10-5 187 601 620 0 TTGGTGAGGTATCCCCGGCG 410533 5-10-5 189 603 622 0 TCTTGGTGAGGTATCCCCGG 410534 5-10-5 191 605 624 0 GATCTTGGTGAGGTATCCCC 410535 5-10-5 192 607 626 0 AGGATCTTGGTGAGGTATCC 410536 5-10-5 243 997 1016 0 GCCACGCCGGCATCCCGGCC 410537 5-10-5 244 998 1017 0 GGCCACGCCGGCATCCCGGC 410538 5-10-5 254 1009 1028 0 CTGGCACCCTTGGCCACGCC 410539 5-10-5 255 1010 1029 0 GCTGGCACCCTTGGCCACGC 410540 5-10-5 356 1572 1591 0 GCTGGTCCTCAGGGAACCAG 410541 5-10-5 376 1849 1868 0 CACCTGGCAATGGCGTAGAC 410542 5-10-5 377 1850 1869 0 GCACCTGGCAATGGCGTAGA 410543 5-10-5 378 1851 1870 0 AGCACCTGGCAATGGCGTAG 410544 5-10-5 379 1852 1871 0 CAGCACCTGGCAATGGCGTA 410545 5-10-5 380 1853 1872 0 GCAGCACCTGGCAATGGCGT 410546 5-10-5 382 1855 1874 0 AGGCAGCACCTGGCAATGGC 410547 5-10-5 384 1857 1876 0 GCAGGCAGCACCTGGCAATG 410548 5-10-5 385 1859 1878 0 TAGCAGGCAGCACCTGGCAA 410549 5-10-5 388 1915 1934 0 GTCCCCATGCTGGCCTCAGC 410550 5-10-5 389 1916 1935 0 GGTCCCCATGCTGGCCTCAG 410551 5-10-5 390 1917 1936 0 GGGTCCCCATGCTGGCCTCA 410552 5-10-5 391 1918 1937 0 CGGGTCCCCATGCTGGCCTC 410553 5-10-5 392 1919 1938 0 ACGGGTCCCCATGCTGGCCT 410554 5-10-5 393 1921 1940 0 ACACGGGTCCCCATGCTGGC 410555 5-10-5 395 1923 1942 0 GGACACGGGTCCCCATGCTG 410556 5-10-5 397 1925 1944 0 GTGGACACGGGTCCCCATGC 410557 5-10-5 399 1927 1946 0 CAGTGGACACGGGTCCCCAT 410558 5-10-5 401 1929 1948 0 GGCAGTGGACACGGGTCCCC 410559 5-10-5 403 1931 1950 0 GTGGCAGTGGACACGGGTCC 410560 5-10-5 404 1932 1951 0 GGTGGCAGTGGACACGGGTC 410561 5-10-5 405 1933 1952 0 TGGTGGCAGTGGACACGGGT 410562 5-10-5 411 2101 2120 0 ACTTTGCATTCCAGACCTGG 410563 5-10-5 413 2103 2122 0 TGACTTTGCATTCCAGACCT 410564 5-10-5 414 2104 2123 0 TTGACTTTGCATTCCAGACC 410565 5-10-5 419 2305 2324 0 CAGATGGCAACGGCTGTCAC 410566 5-10-5 420 2306 2325 0 GCAGATGGCAACGGCTGTCA 410567 5-10-5 421 2307 2326 0 AGCAGATGGCAACGGCTGTC 410568 5-10-5 422 2308 2327 0 CAGCAGATGGCAACGGCTGT 410569 5-10-5 423 2309 2328 0 GCAGCAGATGGCAACGGCTG 410570 5-10-5 424 2311 2330 0 CGGCAGCAGATGGCAACGGC 410571 5-10-5 425 2312 2331 0 CCGGCAGCAGATGGCAACGG 410572 5-10-5 427 2314 2333 0 CTCCGGCAGCAGATGGCAAC 410573 5-10-5 428 2315 2334 0 GCTCCGGCAGCAGATGGCAA 410730 5-10-5 202 657 676 0 CGCCACTCATCTTCACCAGG 410731 5-10-5 207 667 686 0 TCCAGCAGGTCGCCACTCAT 410732 5-10-5 214 791 810 0 GGGCTGGTATTCATCCGCCC 410733 5-10-5 231 886 905 0 AAGTCGGTGACCATGACCCT 410734 5-10-5 279 1219 1238 0 CCGGCAGCGGTGACCAGCAC 410735 5-10-5 291 1320 1339 0 TCCCCAGGGTCACCGGCTGG 410736 5-10-5 303 1356 1375 0 AGAGGTCCACACAGCGGCCA 410737 5-10-5 313 1406 1425 0 GCAGGTGCTGCAGTCGCTGG 410738 5-10-5 324 1482 1501 0 GCTCGGCAGACAGCATCATG 410739 5-10-5 336 1530 1549 0 CAGAGAAGTGGATCAGTCTC 410740 5-10-5 345 1558 1577 0 AACCAGGCCTCATTGATGAC 410741 5-10-5 369 1683 1702 0 CCATCCGTGTAGGCCCCGAG 410742 5-10-5 159 294 313 0 GCCTGGAGCTGACGGTGCCC 410743 5-10-5 170 421 440 0 TCCTCCTCGGAACGCAAGGC 410744 5-10-5 171 446 465 0 GTGCTCGGGTGCTTCGGCCA 410745 5-10-5 172 466 485 0 TGGAAGGTGGCTGTGGTTCC 410746 5-10-5 176 507 526 0 CGTAGGTGCCAGGCAACCTC 410747 5-10-5 177 545 564 0 TGACTGCGAGAGGTGGGTCT 410748 5-10-5 182 591 610 0 ATCCCCGGCGGGCAGCCTGG 410749 5-10-5 197 628 647 0 AGAAGGCCATGGAAGACATG 410750 5-10-5 198 638 657 0 GAAGCCAGGAAGAAGGCCAT 410751 5-10-5 208 685 704 0 GGCAACTTCAAGGCCAGCTC 410752 5-10-5 209 724 743 0 GCAAAGACAGAGGAGTCCTC 410753 5-10-5 215 821 840 0 ATACACCTCCACCAGGCTGC 410754 5-10-5 233 898 917 0 GGCACATTCTCGAAGTCGGT 410755 5-10-5 234 933 952 0 TGGCCTGTCTGTGGAAGCGG 410756 5-10-5 235 960 979 0 GGTGGGTGCCATGACTGTCA 410757 5-10-5 240 985 1004 0 TCCCGGCCGCTGACCACCCC 410758 5-10-5 256 1015 1034 0 CGCATGCTGGCACCCTTGGC 410759 5-10-5 263 1064 1083 0 GGTGCCGCTAACCGTGCCCT 410760 5-10-5 265 1088 1107 0 CCGAATAAACTCCAGGCCTA 410761 5-10-5 271 1119 1138 0 GTGGCCCCACAGGCTGGACC 410762 5-10-5 272 1132 1151 0 AGCAGCACCACCAGTGGCCC 410763 5-10-5 273 1154 1173 0 GCTGTACCCACCCGCCAGGG 410764 5-10-5 274 1200 1219 0 CGACCCCAGCCCTCGCCAGG 410765 5-10-5 286 1273 1292 0 ATGACCTCGGGAGCTGAGGC 410766 5-10-5 287 1283 1302 0 CCCAACTGTGATGACCTCGG 410767 5-10-5 289 1305 1324 0 GCTGGTCTTGGGCATTGGTG 410768 5-10-5 305 1380 1399 0 CAATGATGTCCTCCCCTGGG 410769 5-10-5 318 1425 1444 0 TCCCACTCTGTGACACAAAG 410770 5-10-5 327 1500 1519 0 CGGCCAGGGTGAGCTCCGGC 410771 5-10-5 360 1628 1647 0 ACCTGCCCCATGGGTGCTGG 410772 5-10-5 364 1660 1679 0 GCTGACCATACAGTCCTGCA 410773 5-10-5 387 1905 1924 0 TGGCCTCAGCTGGTGGAGCT 410774 5-10-5 406 1936 1955 0 TGTTGGTGGCAGTGGACACG 410775 5-10-5 407 1962 1981 0 AGCTGCAGCCTGTGAGGACG 410776 5-10-5 408 1990 2009 0 GTGCCAAGGTCCTCCACCTC 410777 5-10-5 409 2010 2029 0 TCAGCACAGGCGGCTTGTGG 410778 5-10-5 410 2040 2059 0 CCACGCACTGGTTGGGCTGA 410779 5-10-5 416 2120 2139 0 CGGGATTCCATGCTCCTTGA 410780 5-10-5 417 2150 2169 0 GCAGGCCACGGTCACCTGCT 410781 5-10-5 418 2187 2206 0 GGAGGGCACTGCAGCCAGTC 410782 5-10-5 429 2325 2344 0 CCAGGTGCCGGCTCCGGCAG 410783 5-10-5 430 2335 2354 0 GAGGCCTGCGCCAGGTGCCG

In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 1. In certain such embodiments, the nucleotide sequences illustrated in Table 2 have a 3-14-3 gap-widened motif. Table 4 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 1, having a 3-14-3 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 4 Gapmer antisense compounds having a 3-14-3 motif targeted to SEQ ID NO: 1 5′ Target Site 3′ Target Site SEQ to SEQ ID to SEQ ID Isis No Motif ID NO NO: 1 NO: 1 Mismatches Sequence (5′-3′) 399871 3-14-3 4 135 154 0 GCGCGGAATCCTGGCTGGGA 399872 3-14-3 5 242 261 0 GAGGAGACCTAGAGGCCGTG 399873 3-14-3 6 300 319 0 AGGACCGCCTGGAGCTGACG 399874 3-14-3 7 410 429 0 ACGCAAGGCTAGCACCAGCT 399949 3-14-3 8 417 436 0 CCTCGGAACGCAAGGCTAGC 399875 3-14-3 9 480 499 0 CCTTGGCGCAGCGGTGGAAG 399876 3-14-3 10 561 580 0 GGCGGGCAGTGCGCTCTGAC 399877 3-14-3 11 600 619 0 TGGTGAGGTATCCCCGGCGG 399950 3-14-3 12 606 625 0 GGATCTTGGTGAGGTATCCC 399951 3-14-3 13 615 634 0 AGACATGCAGGATCTTGGTG 399878 3-14-3 14 620 639 0 ATGGAAGACATGCAGGATCT 399879 3-14-3 15 646 665 0 TTCACCAGGAAGCCAGGAAG 399952 3-14-3 16 651 670 0 TCATCTTCACCAGGAAGCCA 399880 3-14-3 17 705 724 0 CCTCGATGTAGTCGACATGG 399881 3-14-3 18 785 804 0 GTATTCATCCGCCCGGTACC 399882 3-14-3 19 835 854 0 CTGGTGTCTAGGAGATACAC 399953 3-14-3 20 840 859 0 GTATGCTGGTGTCTAGGAGA 399883 3-14-3 21 860 879 0 GATTTCCCGGTGGTCACTCT 399954 3-14-3 22 866 885 0 GCCCTCGATTTCCCGGTGGT 399955 3-14-3 23 880 899 0 GTGACCATGACCCTGCCCTC 399884 3-14-3 24 890 909 0 CTCGAAGTCGGTGACCATGA 399885 3-14-3 25 923 942 0 GTGGAAGCGGGTCCCGTCCT 399886 3-14-3 26 970 989 0 ACCCCTGCCAGGTGGGTGCC 399956 3-14-3 27 975 994 0 TGACCACCCCTGCCAGGTGG 399887 3-14-3 28 1004 1023 0 ACCCTTGGCCACGCCGGCAT 399888 3-14-3 29 1040 1059 0 TTGGCAGTTGAGCACGCGCA 399957 3-14-3 30 1045 1064 0 TTCCCTTGGCAGTTGAGCAC 399889 3-14-3 31 1077 1096 0 CCAGGCCTATGAGGGTGCCG 399890 3-14-3 32 1098 1117 0 GCTGGCTTTTCCGAATAAAC 399891 3-14-3 33 1210 1229 0 GTGACCAGCACGACCCCAGC 399892 3-14-3 149 1297 1316 0 TGGGCATTGGTGGCCCCAAC 399893 3-14-3 34 1326 1345 0 CCAAAGTCCCCAGGGTCACC 399894 3-14-3 128 1330 1349 0 GTCCCCAAAGTCCCCAGGGT 399958 3-14-3 35 1335 1354 0 AGTTGGTCCCCAAAGTCCCC 399895 3-14-3 36 1340 1359 0 GCCAAAGTTGGTCCCCAAAG 399959 3-14-3 37 1352 1371 0 GTCCACACAGCGGCCAAAGT 399896 3-14-3 38 1361 1380 0 GGCAAAGAGGTCCACACAGC 399897 3-14-3 39 1389 1408 0 TGGAGGCACCAATGATGTCC 399960 3-14-3 40 1400 1419 0 GCTGCAGTCGCTGGAGGCAC 399961 3-14-3 41 1411 1430 0 ACAAAGCAGGTGCTGCAGTC 399898 3-14-3 101 1465 1484 0 ATGGCTGCAATGCCAGCCAC 399962 3-14-3 42 1470 1489 0 GCATCATGGCTGCAATGCCA 399963 3-14-3 43 1478 1497 0 GGCAGACAGCATCATGGCTG 399964 3-14-3 44 1526 1545 0 GAAGTGGATCAGTCTCTGCC 399899 3-14-3 45 1534 1553 0 TTGGCAGAGAAGTGGATCAG 399965 3-14-3 46 1539 1558 0 CATCTTTGGCAGAGAAGTGG 399966 3-14-3 47 1545 1564 0 TGATGACATCTTTGGCAGAG 399967 3-14-3 48 1552 1571 0 GCCTCATTGATGACATCTTT 399968 3-14-3 49 1564 1583 0 TCAGGGAACCAGGCCTCATT 399900 3-14-3 50 1569 1588 0 GGTCCTCAGGGAACCAGGCC 399901 3-14-3 87 1576 1595 0 ACCCGCTGGTCCTCAGGGAA 399969 3-14-3 51 1583 1602 0 GGTCAGTACCCGCTGGTCCT 399902 3-14-3 119 1605 1624 0 GCAGGGCGGCCACCAGGTTG 399903 3-14-3 52 1640 1659 0 AAACAGCTGCCAACCTGCCC 399970 3-14-3 53 1645 1664 0 CTGCAAAACAGCTGCCAACC 399904 3-14-3 54 1675 1694 0 GTAGGCCCCGAGTGTGCTGA 399971 3-14-3 55 1740 1759 0 AGAAACTGGAGCAGCTCAGC 399972 3-14-3 56 1746 1765 0 TCCTGGAGAAACTGGAGCAG 399905 3-14-3 57 1812 1831 0 CGTTGTGGGCCCGGCAGACC 399906 3-14-3 58 1858 1877 0 AGCAGGCAGCACCTGGCAAT 399907 3-14-3 59 1920 1939 0 CACGGGTCCCCATGCTGGCC 399908 3-14-3 60 2100 2119 0 CTTTGCATTCCAGACCTGGG 399973 3-14-3 61 2105 2124 0 CTTGACTTTGCATTCCAGAC 399909 3-14-3 62 2310 2329 0 GGCAGCAGATGGCAACGGCT 399910 3-14-3 154 2410 2429 0 TTTTAAAGCTCAGCCCCAGC 399974 3-14-3 63 2415 2434 0 AACCATTTTAAAGCTCAGCC 399911 3-14-3 64 2504 2523 0 TCAAGGGCCAGGCCAGCAGC 399975 3-14-3 65 2509 2528 0 CCCACTCAAGGGCCAGGCCA 399976 3-14-3 122 2582 2601 0 GGAGGGAGCTTCCTGGCACC 399912 3-14-3 66 2597 2616 0 ATGCCCCACAGTGAGGGAGG 399913 3-14-3 67 2606 2625 0 AATGGTGAAATGCCCCACAG 399914 3-14-3 153 2649 2668 0 TTGGGAGCAGCTGGCAGCAC 399915 3-14-3 68 2750 2769 0 CATGGGAAGAATCCTGCCTC 399916 3-14-3 69 2832 2851 0 GGAGATGAGGGCCATCAGCA 399917 3-14-3 70 2900 2919 0 TAGATGCCATCCAGAAAGCT 399977 3-14-3 71 2906 2925 0 TCTGGCTAGATGCCATCCAG 399918 3-14-3 72 2983 3002 0 GGCATAGAGCAGAGTAAAGG 399978 3-14-3 73 2988 3007 0 AGCCTGGCATAGAGCAGAGT 399979 3-14-3 135 2994 3013 0 TAGCACAGCCTGGCATAGAG 399919 3-14-3 112 3227 3246 0 GAAGAGGCTTGGCTTCAGAG 399980 3-14-3 74 3233 3252 0 AAGTAAGAAGAGGCTTGGCT 399920 3-14-3 75 3437 3456 0 GCTCAAGGAGGGACAGTTGT 399921 3-14-3 76 3472 3491 0 AAAGATAAATGTCTGCTTGC 399981 3-14-3 77 3477 3496 0 ACCCAAAAGATAAATGTCTG 399922 3-14-3 78 3543 3562 0 TCTTCAAGTTACAAAAGCAA 399982 3-14-3 99 3550 3569 0 ATAAATATCTTCAAGTTACA 410574 3-14-3 184 597 616 0 TGAGGTATCCCCGGCGGGCA 410575 3-14-3 185 598 617 0 GTGAGGTATCCCCGGCGGGC 410576 3-14-3 186 599 618 0 GGTGAGGTATCCCCGGCGGG 410577 3-14-3 187 601 620 0 TTGGTGAGGTATCCCCGGCG 410578 3-14-3 188 602 621 0 CTTGGTGAGGTATCCCCGGC 410579 3-14-3 189 603 622 0 TCTTGGTGAGGTATCCCCGG 410580 3-14-3 190 604 623 0 ATCTTGGTGAGGTATCCCCG 410581 3-14-3 191 605 624 0 GATCTTGGTGAGGTATCCCC 410582 3-14-3 192 607 626 0 AGGATCTTGGTGAGGTATCC 405604 3-14-3 236 963 982 0 CCAGGTGGGTGCCATGACTG 410583 3-14-3 243 997 1016 0 GCCACGCCGGCATCCCGGCC 410584 3-14-3 244 998 1017 0 GGCCACGCCGGCATCCCGGC 410585 3-14-3 245 999 1018 0 TGGCCACGCCGGCATCCCGG 410586 3-14-3 246 1000 1019 0 TTGGCCACGCCGGCATCCCG 410587 3-14-3 247 1001 1020 0 CTTGGCCACGCCGGCATCCC 410588 3-14-3 248 1002 1021 0 CCTTGGCCACGCCGGCATCC 410589 3-14-3 249 1003 1022 0 CCCTTGGCCACGCCGGCATC 410590 3-14-3 250 1005 1024 0 CACCCTTGGCCACGCCGGCA 410591 3-14-3 251 1006 1025 0 GCACCCTTGGCCACGCCGGC 410592 3-14-3 252 1007 1026 0 GGCACCCTTGGCCACGCCGG 410593 3-14-3 253 1008 1027 0 TGGCACCCTTGGCCACGCCG 410594 3-14-3 254 1009 1028 0 CTGGCACCCTTGGCCACGCC 410595 3-14-3 255 1010 1029 0 GCTGGCACCCTTGGCCACGC 410596 3-14-3 348 1562 1581 0 AGGGAACCAGGCCTCATTGA 410597 3-14-3 349 1563 1582 0 CAGGGAACCAGGCCTCATTG 410598 3-14-3 350 1565 1584 0 CTCAGGGAACCAGGCCTCAT 410599 3-14-3 351 1566 1585 0 CCTCAGGGAACCAGGCCTCA 410600 3-14-3 352 1567 1586 0 TCCTCAGGGAACCAGGCCTC 410601 3-14-3 353 1568 1587 0 GTCCTCAGGGAACCAGGCCT 410602 3-14-3 354 1570 1589 0 TGGTCCTCAGGGAACCAGGC 410603 3-14-3 355 1571 1590 0 CTGGTCCTCAGGGAACCAGG 410604 3-14-3 356 1572 1591 0 GCTGGTCCTCAGGGAACCAG 405641 3-14-3 373 1737 1756 0 AACTGGAGCAGCTCAGCAGC 410605 3-14-3 376 1849 1868 0 CACCTGGCAATGGCGTAGAC 410606 3-14-3 377 1850 1869 0 GCACCTGGCAATGGCGTAGA 410607 3-14-3 378 1851 1870 0 AGCACCTGGCAATGGCGTAG 410608 3-14-3 379 1852 1871 0 CAGCACCTGGCAATGGCGTA 410609 3-14-3 380 1853 1872 0 GCAGCACCTGGCAATGGCGT 410610 3-14-3 381 1854 1873 0 GGCAGCACCTGGCAATGGCG 410611 3-14-3 382 1855 1874 0 AGGCAGCACCTGGCAATGGC 410612 3-14-3 383 1856 1875 0 CAGGCAGCACCTGGCAATGG 410613 3-14-3 384 1857 1876 0 GCAGGCAGCACCTGGCAATG 410614 3-14-3 385 1859 1878 0 TAGCAGGCAGCACCTGGCAA 410615 3-14-3 388 1915 1934 0 GTCCCCATGCTGGCCTCAGC 410616 3-14-3 389 1916 1935 0 GGTCCCCATGCTGGCCTCAG 410617 3-14-3 390 1917 1936 0 GGGTCCCCATGCTGGCCTCA 410618 3-14-3 391 1918 1937 0 CGGGTCCCCATGCTGGCCTC 410619 3-14-3 392 1919 1938 0 ACGGGTCCCCATGCTGGCCT 410620 3-14-3 393 1921 1940 0 ACACGGGTCCCCATGCTGGC 410621 3-14-3 394 1922 1941 0 GACACGGGTCCCCATGCTGG 410622 3-14-3 395 1923 1942 0 GGACACGGGTCCCCATGCTG 410623 3-14-3 396 1924 1943 0 TGGACACGGGTCCCCATGCT 410624 3-14-3 397 1925 1944 0 GTGGACACGGGTCCCCATGC 410625 3-14-3 398 1926 1945 0 AGTGGACACGGGTCCCCATG 410626 3-14-3 399 1927 1946 0 CAGTGGACACGGGTCCCCAT 410627 3-14-3 400 1928 1947 0 GCAGTGGACACGGGTCCCCA 410628 3-14-3 401 1929 1948 0 GGCAGTGGACACGGGTCCCC 410629 3-14-3 402 1930 1949 0 TGGCAGTGGACACGGGTCCC 410630 3-14-3 403 1931 1950 0 GTGGCAGTGGACACGGGTCC 410631 3-14-3 404 1932 1951 0 GGTGGCAGTGGACACGGGTC 410632 3-14-3 405 1933 1952 0 TGGTGGCAGTGGACACGGGT 410633 3-14-3 411 2101 2120 0 ACTTTGCATTCCAGACCTGG 410634 3-14-3 412 2102 2121 0 GACTTTGCATTCCAGACCTG 410635 3-14-3 413 2103 2122 0 TGACTTTGCATTCCAGACCT 410636 3-14-3 414 2104 2123 0 TTGACTTTGCATTCCAGACC 410637 3-14-3 419 2305 2324 0 CAGATGGCAACGGCTGTCAC 410638 3-14-3 420 2306 2325 0 GCAGATGGCAACGGCTGTCA 410639 3-14-3 421 2307 2326 0 AGCAGATGGCAACGGCTGTC 410640 3-14-3 422 2308 2327 0 CAGCAGATGGCAACGGCTGT 410641 3-14-3 423 2309 2328 0 GCAGCAGATGGCAACGGCTG 410642 3-14-3 424 2311 2330 0 CGGCAGCAGATGGCAACGGC 410643 3-14-3 425 2312 2331 0 CCGGCAGCAGATGGCAACGG 410644 3-14-3 426 2313 2332 0 TCCGGCAGCAGATGGCAACG 410645 3-14-3 427 2314 2333 0 CTCCGGCAGCAGATGGCAAC 410646 3-14-3 428 2315 2334 0 GCTCCGGCAGCAGATGGCAA

In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 1. In certain such embodiments, the nucleotide sequences illustrated in Table 2 have a 2-13-5 gap-widened motif. Table 5 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 1, having a 2-13-5 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 5 Gapmer antisense compounds having a 2-13-5 motif targeted to SEQ ID NO: 1 5′ start site SEQ to SEQ ID 3′ Start Site to ISIS No. Motif ID NO NO: 1 SEQ ID NO: 1 Mismatches Sequences (5′ to 3′) 410647 2-13-5 184 597 616 0 TGAGGTATCCCCGGCGGGCA 410648 2-13-5 185 598 617 0 GTGAGGTATCCCCGGCGGGC 410649 2-13-5 186 599 618 0 GGTGAGGTATCCCCGGCGGG 410650 2-13-5 11 600 619 0 TGGTGAGGTATCCCCGGCGG 410651 2-13-5 187 601 620 0 TTGGTGAGGTATCCCCGGCG 410652 2-13-5 188 602 621 0 CTTGGTGAGGTATCCCCGGC 410653 2-13-5 189 603 622 0 TCTTGGTGAGGTATCCCCGG 410654 2-13-5 190 604 623 0 ATCTTGGTGAGGTATCCCCG 410655 2-13-5 191 605 624 0 GATCTTGGTGAGGTATCCCC 410656 2-13-5 12 606 625 0 GGATCTTGGTGAGGTATCCC 410657 2-13-5 192 607 626 0 AGGATCTTGGTGAGGTATCC 410658 2-13-5 243 997 1016 0 GCCACGCCGGCATCCCGGCC 410659 2-13-5 244 998 1017 0 GGCCACGCCGGCATCCCGGC 410660 2-13-5 245 999 1018 0 TGGCCACGCCGGCATCCCGG 410661 2-13-5 246 1000 1019 0 TTGGCCACGCCGGCATCCCG 410662 2-13-5 247 1001 1020 0 CTTGGCCACGCCGGCATCCC 410663 2-13-5 248 1002 1021 0 CCTTGGCCACGCCGGCATCC 410664 2-13-5 249 1003 1022 0 CCCTTGGCCACGCCGGCATC 410665 2-13-5 28 1004 1023 0 ACCCTTGGCCACGCCGGCAT 410666 2-13-5 250 1005 1024 0 CACCCTTGGCCACGCCGGCA 410667 2-13-5 251 1006 1025 0 GCACCCTTGGCCACGCCGGC 410668 2-13-5 252 1007 1026 0 GGCACCCTTGGCCACGCCGG 410669 2-13-5 253 1008 1027 0 TGGCACCCTTGGCCACGCCG 410670 2-13-5 254 1009 1028 0 CTGGCACCCTTGGCCACGCC 410671 2-13-5 255 1010 1029 0 GCTGGCACCCTTGGCCACGC 410672 2-13-5 348 1562 1581 0 AGGGAACCAGGCCTCATTGA 410673 2-13-5 349 1563 1582 0 CAGGGAACCAGGCCTCATTG 410674 2-13-5 49 1564 1583 0 TCAGGGAACCAGGCCTCATT 410675 2-13-5 350 1565 1584 0 CTCAGGGAACCAGGCCTCAT 410676 2-13-5 351 1566 1585 0 CCTCAGGGAACCAGGCCTCA 410677 2-13-5 352 1567 1586 0 TCCTCAGGGAACCAGGCCTC 410678 2-13-5 353 1568 1587 0 GTCCTCAGGGAACCAGGCCT 410679 2-13-5 50 1569 1588 0 GGTCCTCAGGGAACCAGGCC 410680 2-13-5 354 1570 1589 0 TGGTCCTCAGGGAACCAGGC 410681 2-13-5 355 1571 1590 0 CTGGTCCTCAGGGAACCAGG 410682 2-13-5 356 1572 1591 0 GCTGGTCCTCAGGGAACCAG 410683 2-13-5 376 1849 1868 0 CACCTGGCAATGGCGTAGAC 410684 2-13-5 377 1850 1869 0 GCACCTGGCAATGGCGTAGA 410685 2-13-5 378 1851 1870 0 AGCACCTGGCAATGGCGTAG 410686 2-13-5 379 1852 1871 0 CAGCACCTGGCAATGGCGTA 410687 2-13-5 380 1853 1872 0 GCAGCACCTGGCAATGGCGT 410688 2-13-5 381 1854 1873 0 GGCAGCACCTGGCAATGGCG 410689 2-13-5 382 1855 1874 0 AGGCAGCACCTGGCAATGGC 410690 2-13-5 383 1856 1875 0 CAGGCAGCACCTGGCAATGG 410691 2-13-5 384 1857 1876 0 GCAGGCAGCACCTGGCAATG 410692 2-13-5 58 1858 1877 0 AGCAGGCAGCACCTGGCAAT 410693 2-13-5 385 1859 1878 0 TAGCAGGCAGCACCTGGCAA 410694 2-13-5 388 1915 1934 0 GTCCCCATGCTGGCCTCAGC 410695 2-13-5 389 1916 1935 0 GGTCCCCATGCTGGCCTCAG 410696 2-13-5 390 1917 1936 0 GGGTCCCCATGCTGGCCTCA 410697 2-13-5 391 1918 1937 0 CGGGTCCCCATGCTGGCCTC 410698 2-13-5 392 1919 1938 0 ACGGGTCCCCATGCTGGCCT 410699 2-13-5 59 1920 1939 0 CACGGGTCCCCATGCTGGCC 410700 2-13-5 393 1921 1940 0 ACACGGGTCCCCATGCTGGC 410701 2-13-5 394 1922 1941 0 GACACGGGTCCCCATGCTGG 410702 2-13-5 395 1923 1942 0 GGACACGGGTCCCCATGCTG 410703 2-13-5 396 1924 1943 0 TGGACACGGGTCCCCATGCT 410704 2-13-5 397 1925 1944 0 GTGGACACGGGTCCCCATGC 410705 2-13-5 398 1926 1945 0 AGTGGACACGGGTCCCCATG 410706 2-13-5 399 1927 1946 0 CAGTGGACACGGGTCCCCAT 410707 2-13-5 400 1928 1947 0 GCAGTGGACACGGGTCCCCA 410708 2-13-5 401 1929 1948 0 GGCAGTGGACACGGGTCCCC 410709 2-13-5 402 1930 1949 0 TGGCAGTGGACACGGGTCCC 410710 2-13-5 403 1931 1950 0 GTGGCAGTGGACACGGGTCC 410711 2-13-5 404 1932 1951 0 GGTGGCAGTGGACACGGGTC 410712 2-13-5 405 1933 1952 0 TGGTGGCAGTGGACACGGGT 410713 2-13-5 60 2100 2119 0 CTTTGCATTCCAGACCTGGG 410714 2-13-5 411 2101 2120 0 ACTTTGCATTCCAGACCTGG 410715 2-13-5 412 2102 2121 0 GACTTTGCATTCCAGACCTG 410716 2-13-5 413 2103 2122 0 TGACTTTGCATTCCAGACCT 410717 2-13-5 414 2104 2123 0 TTGACTTTGCATTCCAGACC 410718 2-13-5 61 2105 2124 0 CTTGACTTTGCATTCCAGAC 410719 2-13-5 419 2305 2324 0 CAGATGGCAACGGCTGTCAC 410720 2-13-5 420 2306 2325 0 GCAGATGGCAACGGCTGTCA 410721 2-13-5 421 2307 2326 0 AGCAGATGGCAACGGCTGTC 410722 2-13-5 422 2308 2327 0 CAGCAGATGGCAACGGCTGT 410723 2-13-5 423 2309 2328 0 GCAGCAGATGGCAACGGCTG 410724 2-13-5 62 2310 2329 0 GGCAGCAGATGGCAACGGCT 410725 2-13-5 424 2311 2330 0 CGGCAGCAGATGGCAACGGC 410726 2-13-5 425 2312 2331 0 CCGGCAGCAGATGGCAACGG 410727 2-13-5 426 2313 2332 0 TCCGGCAGCAGATGGCAACG 410728 2-13-5 427 2314 2333 0 CTCCGGCAGCAGATGGCAAC 410729 2-13-5 428 2315 2334 0 GCTCCGGCAGCAGATGGCAA

In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 1. In certain such embodiments, the nucleotide sequences illustrated in Table 2 have a 3-13-4 gap-widened motif. Table 6 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 1, having a 3-13-4 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 6 Gapmer antisense compounds having a 3-13-4 motif targeted to SEQ ID NO: 1 SEQ 5′ 3′ ID start Start Mis- Oligo Motif NO: site Site matches OligoSeq 405526 3-13-4 236 963 982 0 CCAGGTGGGTGCC ATGACTG 405557 3-13-4 50 1569 1588 0 GGTCCTCAGGGAA CCAGGCC 405564 3-13-4 373 1737 1756 0 AACTGGAGCAGC TCAGCAGC

The following embodiments set forth target regions of PCSK9 nucleic acids. Also illustrated are examples of antisense compounds targeted to the target regions. It is understood that the sequence set forth in each SEQ ID NO is independent of any modification to a sugar moiety, an internucleoside linkage, or a nucleobase. As such, antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an internucleoside linkage, or a nucleobase. Antisense compounds described by Isis Number (Isis No) indicate a combination of nucleobase sequence and motif.

In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 1: 294-317, 406-440, 406-526, 410-436, 410-499, 446-526, 545-581, 591-619, 591-704, 591-743, 595-622, 600-626, 600-639, 600-670, 601-628, 602-628, 603-630, 611-636, 620-647, 638-665, 648-674, 657-684, 705-743, 782-810, 821-859, 835-859, 835-917, 835-942, 860-887, 860-899, 860-909, 860-917, 869-895, 878-905, 888-909, 923-952, 960-1034, 960-1173, 960-986, 967-991, 970-1023, 970-1064, 970-1117, 970-996, 977-1004, 985-1011, 989-1016, 992-1019, 997-1024, 997-1024, 998-1025, 999-1026, 1000-1027, 1001-1028, 1002-1021, 1002-1029, 1003-1029, 1004-1029, 1005-1029, 1006-1029, 1007-1034, 1036-1061, 1045-1072, 1076-1096, 1088-1115, 1098-1123, 1200-1251, 1210-1237, 1219-1245, 1228-1251, 1273-1444, 1295-1316, 1318-1345, 1328-1354, 1337-1361, 1344-1371, 1354-1377, 1380-1406, 1389-1416, 1400-1426, 1409-1434, 1465-1491, 1465-1602, 1474-1499, 1482-1519, 1513-1540, 1523-1549, 1526-1602, 1526-1624, 1532-1558, 1541-1568, 1552-1579, 1560-1587, 1561-1589, 1564-1591, 1565-1592, 1566-1592, 1567-1592, 1570-1597, 1571-1599, 1605-1706, 1628-1706, 1640-1666, 1672-1698, 1681-1706, 1735-1761, 1735-1765, 1740-1765, 1849-1876, 1849-1879, 1850-1877, 1851-1877, 1852-1878, 1852-1879, 1853-1879, 1854-1879, 1905-1955, 1915-1942, 1916-1943, 1917-1944, 1918-1945, 1919-1946, 1920-1939, 1920-1947, 1921-1948, 1922-1949, 1923-1950, 1924-1951, 1925-1952, 1926-1952, 1927-1952, 1928-1955, 1962-2059, 2040-2126, 2100-2126, 2100-2139, 2100-2206, 2101-2126, 2305-2332, 2305-2354, 2306-2333, 2307-2334, 2308-2334, 2309-2334, 2310-2334, 2410-2434, 2504-2528, 2509-2528, 2582-2625, 2606-2668, 2828-2855, 2832-2851, 2900-2927, 2900-2929, 2902-2927, 2983-3007, 2983-3013, 3227-3252, 3227-3456, 3472-3496, or 3543-3569.

In certain embodiments, a target region is nucleotides 294-317 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 294-317 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 159 or 160. In certain such embodiments, an antisense compound targeted to nucleotides 294-317 of SEQ ID NO: 1 is selected from ISIS NOs: 410742 or 405999.

In certain embodiments, a target region is nucleotides 406-440 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 406-440 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 7, 8, 162, 163, 164, 165, 166, 167, 168, 169 or 170. In certain such embodiments, an antisense compound targeted to nucleotides 406-440 of SEQ ID NO: 1 is selected from ISIS NOs: 405861, 405862, 405863, 405864, 395152, 399874, 405865, 405866, 405867, 405868, 399793, 399949, or 410743.

In certain embodiments, a target region is nucleotides 406-526 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 406-526 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 7, 8, 9, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, or 176. In certain such embodiments, an antisense compound targeted to nucleotides 406-526 of SEQ ID NO: 1 is selected from ISIS NOs: 405861, 405862, 405863, 405864, 395152, 399874, 405865, 405866, 405867, 405868, 399793, 399950, 410743, 410744, 410745, 395153, 399875, 406000, 406001, 406002 or 410746.

In certain embodiments, a target region is nucleotides 410-436 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 410-436 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 7, 8, 166, 167, 168, 169 or 169. In certain such embodiments, an antisense compound targeted to nucleotides 410-436 of SEQ ID NO: 1 is selected from ISIS NOs: 395152, 399874, 405865, 405866, 405867, 405868, or 399793.

In certain embodiments, a target region is nucleotides 410-499 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 410-499 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 7, 8, 9, 166, 167, 169, 170, 171 or 172. In certain such embodiments, an antisense compound targeted to nucleotides 410-499 of SEQ ID NO: 1 is selected from ISIS NOs: 395152, 399874, 405865, 405866, 405867, 405868, 399793, 399949, 410743, 410744, 410745, 395153, or 399875.

In certain embodiments, a target region is nucleotides 446-526 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 446-526 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 9, 171, 172, 173, 174, 175 or 176. In certain such embodiments, an antisense compound targeted to nucleotides 446-526 of SEQ ID NO: 1 is selected from ISIS NOs: 410744, 410745, 395153, 399875, 406000, 406001, 406002, or 410746.

In certain embodiments, a target region is nucleotides 545-581 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 545-581 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 10, 177, 178, 179, 180 or 181. In certain such embodiments, an antisense compound targeted to nucleotides 545-581 of SEQ ID NO: 1 is selected from ISIS NOs: 410747, 406003, 406004, 406005, 395154, 399876, or 406006.

In certain embodiments, a target region is nucleotides 591-619 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 591-619 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 182, 183, 184, 185 or 186. In certain such embodiments, an antisense compound targeted to nucleotides 591-619 of SEQ ID NO: 1 is selected from ISIS NOs: 410748, 406007, 410529, 410574, 410647, 410530, 410575, 410648, 410531, 410576, 410649, 395155, 399877, or 410650.

In certain embodiments, a target region is nucleotides 591-704 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 591-704 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 14, 15, 16, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, or 208. In certain such embodiments, an antisense compound targeted to nucleotides 591-704 of SEQ ID NO: 1 is selected from ISIS NOs: 410748, 406007, 410529, 410574, 410647, 410530, 410575, 410648, 410531, 410576, 410649, 395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, or 410751.

In certain embodiments, a target region is nucleotides 591-743 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 591-743 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 14, 15, 16, 17, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, or 209. In certain such embodiments, an antisense compound targeted to nucleotides 591-743 of SEQ ID NO: 1 is selected from ISIS NOs: 410748, 406007, 410529, 410574, 410647, 410530, 410575, 410648, 410531, 410576, 410649, 395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, or 410752.

In certain embodiments, a target region is nucleotides 595-622 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 595-622 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 183, 184, 185, 186, 187, 188, or 189. In certain such embodiments, an antisense compound targeted to nucleotides 595-622 of SEQ ID NO: 1 is selected from ISIS NOs: 406007, 410529, 410574, 410647, 410530, 410575, 410648, 410531, 410576, 410649, 395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, or 410653.

In certain embodiments, a target region is nucleotides 600-626 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 600-626 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 87, 188, 189, 190, 191, or 192. In certain such embodiments, an antisense compound targeted to nucleotides 600-626 of SEQ ID NO: 1 is selected from ISIS NOs: 395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, or 410657.

In certain embodiments, a target region is nucleotides 600-639 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 600-639 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 14, 187, 188, 189, 190, 191, 192, 193, 194, 195, or 196. In certain such embodiments, an antisense compound targeted to nucleotides 600-639 of SEQ ID NO: 1 is selected from ISIS NOs: 395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, 410657, 406010, 406011, 406012, 399795, 399951, 406013, 395156, or 399878.

In certain embodiments, a target region is nucleotides 600-670 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 600-670 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 14, 15, 16, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199. In certain such embodiments, an antisense compound targeted to nucleotides 600-670 of SEQ ID NO: 1 is selected from ISIS NOs: 395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, 410657, 406010, 406011, 406012, 399795, 399951, 406013, 395156, 399878, or 399952.

In certain embodiments, a target region is nucleotides 601-628 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 601-628 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 187, 188, 189, 190, 191, 192 or 193. In certain such embodiments, an antisense compound targeted to nucleotides 601-628 of SEQ ID NO: 1 is selected from ISIS NOs: 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, 410657, or 406010.

In certain embodiments, a target region is nucleotides 602-628 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 602-628 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 188, 189, 190, 191, 192 or 193. In certain such embodiments, an antisense compound targeted to nucleotides 602-628 of SEQ ID NO: 1 is selected from ISIS NOs: 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, 410657, or 406010.

In certain embodiments, a target region is nucleotides 603-630 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 603-630 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 189, 190, 191, 192, 193 or 194. In certain such embodiments, an antisense compound targeted to nucleotides 603-630 of SEQ ID NO: 1 is selected from ISIS NOs: 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, 410657, 406010, or 406011.

In certain embodiments, a target region is nucleotides 611-636 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 611-636 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 13, 194, 195 or 196. In certain such embodiments, an antisense compound targeted to nucleotides 611-636 of SEQ ID NO: 1 is selected from ISIS NOs: 406011, 406012, 399795, 399951, or 406013.

In certain embodiments, a target region is nucleotides 620-647 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 620-647 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 14 or 197. In certain such embodiments, an antisense compound targeted to nucleotides 620-647 of SEQ ID NO: 1 is selected from ISIS NOs: 395156, 399878, or 410749.

In certain embodiments, a target region is nucleotides 638-665 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 638-665 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 15 or 198. In certain such embodiments, an antisense compound targeted to nucleotides 638-665 of SEQ ID NO: 1 is selected from ISIS NOs: 410750, 395157, or 399879.

In certain embodiments, a target region is nucleotides 648-674 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 648-674 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 16, 199, 200, or 201. In certain such embodiments, an antisense compound targeted to nucleotides 648-674 of SEQ ID NO: 1 is selected from ISIS NOs: 406014, 399796, 399952, 406015, or 406016.

In certain embodiments, a target region is nucleotides 657-684 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 657-684 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 202, 203, 204, 205, or 206. In certain such embodiments, an antisense compound targeted to nucleotides 657-684 of SEQ ID NO: 1 is selected from ISIS NOs: 410730, 406017, 406018, 406019, or 406020.

In certain embodiments, a target region is nucleotides 705-743 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 705-743 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 17 or 209. In certain such embodiments, an antisense compound targeted to nucleotides 705-743 of SEQ ID NO: 1 is selected from ISIS NOs: 395158, 399880, or 410752.

In certain embodiments, a target region is nucleotides 782-810 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 782-810 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 18, 210, 211, 212, 213, or 214. In certain such embodiments, an antisense compound targeted to nucleotides 782-810 of SEQ ID NO:1 is selected from ISIS NOs: 406021, 406022, 395159, 399881, 406023, 406024, or 410732.

In certain embodiments, a target region is nucleotides 821-859 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 821-859 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20, 215, 216 or 217. In certain such embodiments, an antisense compound targeted to nucleotides 821-859 of SEQ ID NO: 1 is selected from ISIS NOs: 410753, 406025, 395160, 399882, 406026, 399797, or 399953.

In certain embodiments, a target region is nucleotides 835-859 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 835-859 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20 or 217. In certain such embodiments, an antisense compound targeted to nucleotides 835-859 of SEQ ID NO: 1 is selected from ISIS NOs: 395160, 399882, 406026, 399797, or 399953.

In certain embodiments, a target region is nucleotides 835-917 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 835-917 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20, 21, 22, 23, 24, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232 or 233. In certain such embodiments, an antisense compound targeted to nucleotides 835-917 of SEQ ID NO: 1 is selected from ISIS NOs: 395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, 399884, or 410754.

In certain embodiments, a target region is nucleotides 835-942 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 835-942 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20, 21, 22, 23, 24, 25, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, or 233. In certain such embodiments, an antisense compound targeted to nucleotides 835-942 of SEQ ID NO: 1 is selected from ISIS NOs: 395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, 399884, 410754, 395163, or 399885.

In certain embodiments, a target region is nucleotides 860-887 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 860-887 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 218, 219, 220, 221, 222 or 223. In certain such embodiments, an antisense compound targeted to nucleotides 860-887 of SEQ ID NO: 1 is selected from ISIS NOs: 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, and 405874.

In certain embodiments, a target region is nucleotides 860-899 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 860-899 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227 or 228. In certain such embodiments, an antisense compound targeted to nucleotides 860-899 of SEQ ID NO: 1 is selected from ISIS NOs: 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, or 399955.

In certain embodiments, a target region is nucleotides 860-909 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 860-909 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 24, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231 or 232. In certain such embodiments, an antisense compound targeted to nucleotides 860-909 of SEQ ID NO: 1 is selected from ISIS NOs: 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, or 399884.

In certain embodiments, a target region is nucleotides 860-917 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 860-917 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 24, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, or 233. In certain such embodiments, an antisense compound targeted to nucleotides 860-917 of SEQ ID NO: 1 is selected from ISIS NOs: 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, 399884, or 410754.

In certain embodiments, a target region is nucleotides 869-895 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 869-895 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 224, 225, 226, or 227. In certain such embodiments, an antisense compound targeted to nucleotides 869-895 of SEQ ID NO: 1 is selected from ISIS NOs: 405875, 405876, 406027, or 406028.

In certain embodiments, a target region is nucleotides 878-905 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 878-905 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 23, 228, 229, 230, or 231. In certain such embodiments, an antisense compound targeted to nucleotides 878-905 of SEQ ID NO: 1 is selected from ISIS NOs: 406029, 399799, 399955, 406030, 406031, or 410733.

In certain embodiments, a target region is nucleotides 888-909 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 888-909 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 24 or 232. In certain such embodiments, an antisense compound targeted to nucleotides 888-909 of SEQ ID NO: 1 is selected from ISIS NOs: 406032, 395162, or 399884.

In certain embodiments, a target region is nucleotides 923-952 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 923-952 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 25 or 234. In certain such embodiments, an antisense compound targeted to nucleotides 923-952 of SEQ ID NO: 1 is selected from ISIS NOs: 395163, 399885, or 410755.

In certain embodiments, a target region is nucleotides 960-1034 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 960-1034 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, or 256. In certain such embodiments, an antisense compound targeted to nucleotides 960-1034 of SEQ ID NO: 1 is selected from ISIS NOs: 410756, 405526, 405604, 406033, 395164, 399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, or 410758.

In certain embodiments, a target region is nucleotides 960-1173 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 960-1173 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, or 273. In certain such embodiments, an antisense compound targeted to nucleotides 960-1173 of SEQ ID NO: 1 is selected from ISIS NOs: 410756, 405526, 405604, 406033, 395164, 399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, or 410763.

In certain embodiments, a target region is nucleotides 960-986 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 960-986 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 235, 236, or 237. In certain such embodiments, an antisense compound targeted to nucleotides 960-986 of SEQ ID NO: 1 is selected from ISIS NOs: 410756, 405526, 405604, or 406033.

In certain embodiments, a target region is nucleotides 967-991 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 967-991 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 237 or 238. In certain such embodiments, an antisense compound targeted to nucleotides 967-991 of SEQ ID NO: 1 is selected from ISIS NOs: 406033 or 406034.

In certain embodiments, a target region is nucleotides 970-1023 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 970-1023 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, or 249. In certain such embodiments, an antisense compound targeted to nucleotides 970-1023 of SEQ ID NO: 1 is selected from ISIS NOs: 395164, 399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, or 410665.

In certain embodiments, a target region is nucleotides 970-1064 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 970-1064 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 29, 30, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 149, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, or 263. In certain such embodiments, an antisense compound targeted to nucleotides 970-1064 of SEQ ID NO: 1 is selected from ISIS NOs: 395164, 399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, or 399957.

In certain embodiments, a target region is nucleotides 970-1117 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 970-1117 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 29, 30, 31, 32, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 149, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, or 266. In certain such embodiments, an antisense compound targeted to nucleotides 970-1117 of SEQ ID NO: 1 is selected from ISIS NOs: 395164, 399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, or 399890.

In certain embodiments, a target region is nucleotides 970-996 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 970-996 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, or 238. In certain such embodiments, an antisense compound targeted to nucleotides 970-996 of SEQ ID NO: 1 is selected from ISIS NOs: 395164, 399886, 406034, 399800, 399956, or 406035.

In certain embodiments, a target region is nucleotides 977-1004 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 977-1004 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 239 or 240. In certain such embodiments, an antisense compound targeted to nucleotides 977-1004 of SEQ ID NO: 1 is selected from ISIS NOs: 406035 or 410757.

In certain embodiments, a target region is nucleotides 985-1011 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 985-1011 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 240, 241, or 242. In certain such embodiments, an antisense compound targeted to nucleotides 985-1011 of SEQ ID NO: 1 is selected from ISIS NOs: 410757, 406036, or 406037.

In certain embodiments, a target region is nucleotides 989-1016 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 989-1016 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 241, 242, or 243. In certain such embodiments, an antisense compound targeted to nucleotides 989-1016 of SEQ ID NO: 1 is selected from ISIS NOs: 406036, 406037, 410536, 410583, or 410658.

In certain embodiments, a target region is nucleotides 992-1019 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 992-1019 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 242, 243, 244, 245, or 246. In certain such embodiments, an antisense compound targeted to nucleotides 992-1019 of SEQ ID NO: 1 is selected from ISIS NOs: 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, or 410661.

In certain embodiments, a target region is nucleotides 997-1024 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 997-1024 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 243, 244, 245, 246, 247, 248, 249, or 250. In certain such embodiments, an antisense compound targeted to nucleotides 997-1024 of SEQ ID NO: 1 is selected from ISIS NOs: 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 409126, 410665, 405881, 410590, or 410666.

In certain embodiments, a target region is nucleotides 997-1024 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 997-1024 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 243, 244, 245, 246, 247, 248, 249, or 250. In certain such embodiments, an antisense compound targeted to nucleotides 997-1024 of SEQ ID NO: 1 is selected from ISIS NOs: 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 409126, 410665, 405881, 410590, or 410666.

In certain embodiments, a target region is nucleotides 998-1025 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 998-1025 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 244, 245, 246, 247, 248, 249, 250, or 251. In certain such embodiments, an antisense compound targeted to nucleotides 998-1025 of SEQ ID NO: 1 is selected from ISIS NOs: 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, or 410667.

In certain embodiments, a target region is nucleotides 999-1026 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 999-1026 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 245, 246, 247, 248, 249, 250, 251, or 252. In certain such embodiments, an antisense compound targeted to nucleotides 999-1026 of SEQ ID NO: 1 is selected from ISIS NOs: 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405833, 410592, or 410668.

In certain embodiments, a target region is nucleotides 1000-1027 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1000-1027 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 246, 247, 248, 249, 250, 251, 252, or 253. In certain such embodiments, an antisense compound targeted to nucleotides 1000-1027 of SEQ ID NO: 1 is selected from ISIS NOs: 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405833, 410592, 410668, 405884, 410593, or 410669.

In certain embodiments, a target region is nucleotides 1001-1028 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1001-1028 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 247, 248, 249, 250, 251, 252, 253, or 254. In certain such embodiments, an antisense compound targeted to nucleotides 1001-1028 of SEQ ID NO: 1 is selected from ISIS NOs: 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405833, 410592, 410668, 405884, 410593, 410669, 410538, 410594, or 410670.

In certain embodiments, a target region is nucleotides 1002-1021 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1002-1021 of SEQ ID NO: 1. In one such embodiment, an antisense compound targeted to nucleotides 1002-1021 of SEQ ID NO: 1 is ISIS NO: 405879.

In certain embodiments, a target region is nucleotides 1002-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1002-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 248, 249, 250, 251, 252, 253, 254, or 255. In certain such embodiments, an antisense compound targeted to nucleotides 1002-1029 of SEQ ID NO: 1 is selected from ISIS NOs: 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405833, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.

In certain embodiments, a target region is nucleotides 1003-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1003-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 249, 250, 251, 252, 253, 254, or 255. In certain such embodiments, an antisense compound targeted to nucleotides 1003-1029 of SEQ ID NO: 1 is selected from ISIS NOs: 405880, 410589, 410664, 395165, 399887, 409126, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.

In certain embodiments, a target region is nucleotides 1004-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1004-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 250, 251, 252, 253, 254, or 255. In certain such embodiments, an antisense compound targeted to nucleotides 1004-1029 of SEQ ID NO: 1 is selected from ISIS NOs: 395165, 399887, 409126, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.

In certain embodiments, a target region is nucleotides 1005-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1005-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NQs: 250, 251, 252, 253, 254, or 255. In certain such embodiments, an antisense compound targeted to nucleotides 1005-1029 of SEQ ID NO: 1 is selected from ISIS NOs: 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.

In certain embodiments, a target region is nucleotides 1006-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1006-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 251, 252, 253, 254, or 255. In certain such embodiments, an antisense compound targeted to nucleotides 1006-1029 of SEQ ID NO: 1 is selected from ISIS NOs: 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.

In certain embodiments, a target region is nucleotides 1007-1034 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1007-1034 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 252, 253, 254, 255, or 256. In certain such embodiments, an antisense compound targeted to nucleotides 1007-1034 of SEQ ID NO: 1 is selected from ISIS NOs: 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, 410671, or 410758.

In certain embodiments, a target region is nucleotides 1036-1061 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1036-1061 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 29, 257, 258, or 259. In certain such embodiments, an antisense compound targeted to nucleotides 1036-1061 of SEQ ID NO: 1 is selected from ISIS NOs: 406039, 406040, 395166, 399888, or 406041.

In certain embodiments, a target region is nucleotides 1045-1072 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1045-1072 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 30, 260, 261, or 262. In certain such embodiments, an antisense compound targeted to nucleotides 1045-1072 of SEQ ID NO: 1 is selected from ISIS NOs: 399801, 399957, 406042, 406043, or 406044.

In certain embodiments, a target region is nucleotides 1076-1096 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1076-1096 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 31 or 264. In certain such embodiments, an antisense compound targeted to nucleotides 1076-1096 of SEQ ID NO: 1 is selected from ISIS NOs: 408642, 395167, or 399889.

In certain embodiments, a target region is nucleotides 1088-1115 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1088-1115 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 265 or 266. In certain such embodiments, an antisense compound targeted to nucleotides 1088-1115 of SEQ ID NO: 1 is selected from ISIS NOs: 410760 or 406045.

In certain embodiments, a target region is nucleotides 1098-1123 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1098-1123 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 32, 267, 268, or 269. In certain such embodiments, an antisense compound targeted to nucleotides 1098-1123 of SEQ ID NO: 1 is selected from ISIS NOs: 395168, 399890, 405909, 405910, or 405911.

In certain embodiments, a target region is nucleotides 1200-1251 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1200-1251 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 33, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, or 285. In certain such embodiments, an antisense compound targeted to nucleotides 1200-1251 of SEQ ID NO: 1 is selected from ISIS NOs: 410764, 395169, 399891, 405913, 405914, 405915, 405916, 410734, 405917, 405918, 405919, 405920, 405921, or 405922.

In certain embodiments, a target region is nucleotides 1210-1237 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1210-1237 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 33, 275, 276, 277, or 278. In certain such embodiments, an antisense compound targeted to nucleotides 1210-1237 of SEQ ID NO: 1 is selected from ISIS NOs: 395169, 399891, 405913, 405914, 405915, or 405916.

In certain embodiments, a target region is nucleotides 1219-1245 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1219-1245 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 279, 280, 281 or 282. In certain such embodiments, an antisense compound targeted to nucleotides 1219-1245 of SEQ ID NO: 1 is selected from ISIS NOs: 410734, 405917, 405918, or 405919.

In certain embodiments, a target region is nucleotides 1228-1251 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1228-1251 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 283, 284, or 285. In certain such embodiments, an antisense compound targeted to nucleotides 1228-1251 of SEQ ID NO:1 is selected from ISIS NOs: 405920, 405921, or 405922.

In certain embodiments, a target region is nucleotides 1273-1444 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1273-1444 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 34, 35, 36, 37, 38, 39, 40, 41, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 316, and 318. In certain such embodiments, an antisense compound targeted to nucleotides 1273-1444 of SEQ ID NO: 1 is selected from ISIS NOs: 410765, 410766, 405923, 395170, 399892, 410767, 405924, 410735, 405925, 405926, 395171, 399893, 405927, 395172, 399894, 405928, 399802, 399958, 405929, 395173, 399895, 405930, 405931, 405932, 405933, 405934, 399803, 399959, 405935, 410736, 405936, 395174, or 410769.

In certain embodiments, a target region is nucleotides 1295-1316 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1295-1316 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 149 or 288. In certain such embodiments, an antisense compound targeted to nucleotides 1295-1316 of SEQ ID NO: 1 is selected from ISIS NOs: 405923, 395170, or 399892.

In certain embodiments, a target region is nucleotides 1318-1345 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1318-1345 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 34, 290, 291, 292 or 293. In certain such embodiments, an antisense compound targeted to nucleotides 1318-1345 of SEQ ID NO: 1 is selected from ISIS NOs: 405924, 410735, 405925, 405926, 395171, or 399893.

In certain embodiments, a target region is nucleotides 1328-1354 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1328-1354 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 35, 128, 294 or 295. In certain such embodiments, an antisense compound targeted to nucleotides 1328-1354 of SEQ ID NO: 1 is selected from ISIS NOs: 405927, 395172, 399894, 405928, 399802, or 399958.

In certain embodiments, a target region is nucleotides 1337-1361 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1337-1361 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 36, 296, or 297. In certain such embodiments, an antisense compound targeted to nucleotides 1337-1361 of SEQ ID NO: 1 is selected from ISIS NOs: 405929, 395173, 399895, or 405930.

In certain embodiments, a target region is nucleotides 1344-1371 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1344-1371 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 37, 298, 299, 300, or 301. In certain such embodiments, an antisense compound targeted to nucleotides 1344-1371 of SEQ ID NO: 1 is selected from ISIS NOs: 405931, 405932, 405933, 405934, 399803, or 399959.

In certain embodiments, a target region is nucleotides 1354-1377 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1354-1377 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 302, 303, or 304. In certain such embodiments, an antisense compound targeted to nucleotides 1354-1377 of SEQ ID NO: 1 is selected from ISIS NOs: 405935, 410736, or 405936.

In certain embodiments, a target region is nucleotides 1380-1406 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1380-1406 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 305 or 306. In certain such embodiments, an antisense compound targeted to nucleotides 1380-1406 of SEQ ID NO: 1 is selected from ISIS NOs: 410768 or 405937.

In certain embodiments, a target region is nucleotides 1389-1416 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1389-1416 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 39, 307, 308, 309 or 310. In certain such embodiments, an antisense compound targeted to nucleotides 1389-1416 of SEQ ID NO: 1 is selected from ISIS NOs: 395175, 399897, 405938, 405939, 405940, or 405941.

In certain embodiments, a target region is nucleotides 1400-1426 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1400-1426 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 40, 311, 312, 313, or 314. In certain such embodiments, an antisense compound targeted to nucleotides 1400-1426 of SEQ ID NO: 1 is selected from ISIS NOs: 399804, 399960, 405942, 405943, 410737, or 405944.

In certain embodiments, a target region is nucleotides 1409-1434 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1409-1434 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 41, 315, 316, or 317. In certain such embodiments, an antisense compound targeted to nucleotides 1409-1434 of SEQ ID NO: 1 is selected from ISIS NOs: 405945, 399805, 399961, 405946, or 405947.

In certain embodiments, a target region is nucleotides 1465-1491 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1465-1491 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 42, 101, 319, or 320. In certain such embodiments, an antisense compound targeted to nucleotides 1465-1491 of SEQ ID NO: 1 is selected from ISIS Nos: 395176, 399898, 405948, 399806, 399962, or 405949.

In certain embodiments, a target region is nucleotides 1465-1602 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1465-1602 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 87, 101, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, or 359. In certain such embodiments, an antisense compound targeted to nucleotides 1465-1602 of SEQ ID NO: 1 is selected from ISIS NOs: 395176, 399898, 405948, 399806, 399962, 405949, 405950, 405951, 399807, 399963, 405952, 410738, 405953, 405954, 410770, 405955, 405956, 405957, 405958, 405959, 405960, 405961, 399808, 399964, 405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, or 399969.

In certain embodiments, a target region is nucleotides 1474-1499 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1474-1499 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 321, 322, or 323. In certain such embodiments, an antisense compound targeted to nucleotides 1474-1499 of SEQ ID NO: 1 is selected from ISIS NOs: 405950, 405951, 399807, 399963, or 405952.

In certain embodiments, a target region is nucleotides 1482-1519 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1482-1519 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 324, 325, 326, or 327. In certain such embodiments, an antisense compound targeted to nucleotides 1482-1519 of SEQ ID NO: 1 is selected from ISIS NOs: 410738, 405953, 405954, or 410770.

In certain embodiments, a target region is nucleotides 1513-1540 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1513-1540 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 328, 329, 330, 331, or 332. In certain such embodiments, an antisense compound targeted to nucleotides 1513-1540 of SEQ ID NO: 1 is selected from ISIS NOs: 405955, 405956, 405957, 405958, or 405959.

In certain embodiments, a target region is nucleotides 1523-1549 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1523-1549 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 333, 334, 335, or 336. In certain such embodiments, an antisense compound targeted to nucleotides 1523-1549 of SEQ ID NO: 1 is selected from ISIS NOs: 405960, 405961, 399808, 399964, 405962, or 410739.

In certain embodiments, a target region is nucleotides 1526-1602 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1526-1602 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 45, 46, 47, 48, 49, 50, 51, 87, 335, 336, 337, 338, 339, 340, 341, 342, 343, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, or 359. In certain such embodiments, an antisense compound targeted to nucleotides 1526-1602 of SEQ ID NO: 1 is selected from ISIS NOs: 399808, 399964, 405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, 405965, 405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969, 405970, 410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, or 399969.

In certain embodiments, a target region is nucleotides 1526-1624 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1526-1624 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 45, 46, 47, 48, 49, 50, 51, 87, 119, 335, 336, 337, 338, 339, 340, 341, 342, 343, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, or 359. In certain such embodiments, an antisense compound targeted to nucleotides 1526-1624 of SEQ ID NO: 1 is selected from ISIS NOs: 399808, 399964, 405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, 405965, 405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969, 405970, 410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, or 399902.

In certain embodiments, a target region is nucleotides 1532-1558 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1532-1558 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 45, 46, 337, or 338. In certain such embodiments, an antisense compound targeted to nucleotides 1532-1558 of SEQ ID NO: 1 is selected from ISIS NOs: 405963, 395177, 399899, 405964, 399809, or 399965.

In certain embodiments, a target region is nucleotides 1541-1568 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1541-1568 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 47, 340, 341, or 342. In certain such embodiments, an antisense compound targeted to nucleotides 1541-1568 of SEQ ID NO: 1 is selected from ISIS NOs: 405965, 405966, 399810, 399966, 405967, or 405968.

In certain embodiments, a target region is nucleotides 1552-1579 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1552-1579 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 48, 343, 344, 345, or 346. In certain such embodiments, an antisense compound targeted to nucleotides 1552-1579 of SEQ ID NO: 1 is selected from ISIS NOs: 399811, 399967, 405969, 405970, 410740, or 405885.

In certain embodiments, a target region is nucleotides 1560-1587 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1560-1587 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 346, 347, 348, 349, 350, 351, 352, or 353. In certain such embodiments, an antisense compound targeted to nucleotides 1560-1587 of SEQ ID NO: 1 is selected from ISIS NOs: 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, or 410678.

In certain embodiments, a target region is nucleotides 1561-1589 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1561-1589 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 346, 347, 348, 349, 350, 351, 352, 353, or 354. In certain such embodiments, an antisense compound targeted to nucleotides 1561-1589 of SEQ ID NO: 1 is selected from ISIS NOs: 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 405557, 410679, 408653, 410602, or 410680.

In certain embodiments, a target region is nucleotides 1564-1591 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1564-1591 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 350, 351, 352, 353, 354, 355, or 356. In certain such embodiments, an antisense compound targeted to nucleotides 1564-1591 of SEQ ID NO: 1 is selected from ISIS NOs: 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 405557, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, or 410682.

In certain embodiments, a target region is nucleotides 1565-1592 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1565-1592 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 50, 350, 351, 352, 353, 354, 355, 356, or 357. In certain such embodiments, an antisense compound targeted to nucleotides 1565-1592 of SEQ ID NO: 1 is selected from ISIS NOs: 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 405557, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.

In certain embodiments, a target region is nucleotides 1566-1592 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1566-1592 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 50, 351, 352, 353, 354, 355, 356, or 357. In certain such embodiments, an antisense compound targeted to nucleotides 1566-1592 of SEQ ID NO: 1 is selected from ISIS NOs: 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 405557, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.

In certain embodiments, a target region is nucleotides 1567-1592 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1567-1592 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 50, 352, 353, 354, 355, 356, or 357. In certain such embodiments, an antisense compound targeted to nucleotides 1567-1592 of SEQ ID NO: 1 is selected from ISIS NOs: 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 405557, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.

In certain embodiments, a target region is nucleotides 1570-1597 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1570-1597 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 354, 355, 356, 357 or 358. In certain such embodiments, an antisense compound targeted to nucleotides 1570-1597 of SEQ ID NO: 1 is selected from ISIS NOs: 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, 405972 or 405973.

In certain embodiments, a target region is nucleotides 1571-1599 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1571-1599 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 87, 356, 357, 358, or 359. In certain such embodiments, an antisense compound targeted to nucleotides 1571-1599 of SEQ ID NO: 1 is selected from ISIS NOs: 405971, 410603, 410681, 410540, 410604, 410682, 405972, 395179, 399901, 405973, or 405974.

In certain embodiments, a target region is nucleotides 1605-1706 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1605-1706 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 52, 53, 54, 119, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, or 371. In certain such embodiments, an antisense compound targeted to nucleotides 1605-1706 of SEQ ID NO: 1 is selected from ISIS NOs: 395180, 399902, 410771, 395181, 399903, 405975, 399814, 399970, 405976, 405977, 410772, 405978, 395182, 399904, 405979, 405980, 405981, 410741, 405982, or 405983.

In certain embodiments, a target region is nucleotides 1628-1706 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1628-1706 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 52, 53, 54, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, or 371. In certain such embodiments, an antisense compound targeted to nucleotides 1628-1706 of SEQ ID NO: 1 is selected from ISIS NOs: 410771, 395181, 399903, 405975, 399814, 399970, 405976, 405977, 410772, 405978, 395182, 399904, 405979, 405980, 405981, 410741, 405982, or 405983.

In certain embodiments, a target region is nucleotides 1640-1666 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1640-1666 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 52, 53, 361, or 362. In certain such embodiments, an antisense compound targeted to nucleotides 1640-1666 of SEQ ID NO:1 is selected from ISIS NOs: 395181, 399903, 405975, 399814, 399970, or 405976.

In certain embodiments, a target region is nucleotides 1672-1698 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1672-1698 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 54, 365, 366, or 367. In certain such embodiments, an antisense compound targeted to nucleotides 1672-1698 of SEQ ID NO: 1 is selected from ISIS NOs: 405978, 395182, 399904, 405979, or 405980.

In certain embodiments, a target region is nucleotides 1681-1706 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1681-1706 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 368, 369, 370, or 371. In certain such embodiments, an antisense compound targeted to nucleotides 1681-1706 of SEQ ID NO: 1 is selected from ISIS NOs: 405981, 410741, 405982, or 405983.

In certain embodiments, a target region is nucleotides 1735-1761 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1735-1761 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 372, 373, or 374. In certain such embodiments, an antisense compound targeted to nucleotides 1735-1761 of SEQ ID NO: 1 is selected from ISIS NOs: 405984, 405564, 405641, 405985, 399815, 399971, or 405986.

In certain embodiments, a target region is nucleotides 1735-1765 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1735-1765 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 56, 372, 373, 374, or 375. In certain such embodiments, an antisense compound targeted to nucleotides 1735-1765 of SEQ ID NO: 1 is selected from ISIS NOs: 405984, 405564, 405641, 405985, 399815, 399971, 405986, 405987, 399816 or 399972.

In certain embodiments, a target region is nucleotides 1740-1765 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1740-1765 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 56, 374, or 375. In certain such embodiments, an antisense compound targeted to nucleotides 1740-1765 of SEQ ID NO: 1 is selected from ISIS NOs: 399815, 399971, 405986, 405987, 399816, or 399972.

In certain embodiments, a target region is nucleotides 1849-1876 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1849-1876 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 376, 377, 378, 379, 380, 381, 382, 383, or 384. In certain such embodiments, an antisense compound targeted to nucleotides 1849-1876 of SEQ ID NO: 1 is selected from ISIS NOs: 410541, 410605, 410683, 410542, 410606, 410684, 410543, 410607, 410685, 410544, 410608, 410686, 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, or 410691.

In certain embodiments, a target region is nucleotides 1849-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1849-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, or 386. In certain such embodiments, an antisense compound targeted to nucleotides 1849-1879 of SEQ ID NO: 1 is selected from ISIS NOs: 410541, 410605, 410683, 410542, 410606, 410684, 410543, 410607, 410685, 410544, 410608, 410686, 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, 410692, 410548, 410614, or 405990.

In certain embodiments, a target region is nucleotides 1850-1877 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1850-1877 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 377, 378, 379, 380, 381, 382, 383 or 384. In certain such embodiments, an antisense compound targeted to nucleotides 1850-1877 of SEQ ID NO: 1 is selected from ISIS NOs: 410542, 410606, 410684, 410543, 410607, 410685, 410544, 410608, 410686, 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, or 410692.

In certain embodiments, a target region is nucleotides 1851-1877 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1851-1877 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 378, 379, 380, 381, 382, 383, or 384. In certain such embodiments, an antisense compound targeted to nucleotides 1851-1877 of SEQ ID NO: 1 is selected from ISIS NOs: 410543, 410607, 410685, 410544, 410608, 410686, 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, or 410692.

In certain embodiments, a target region is nucleotides 1852-1878 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1852-1878 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID Nos: 58, 379, 380, 381, 382, 383, 384, or 385. In certain such embodiments, an antisense compound targeted to nucleotides 1852-1878 of SEQ ID NO: 1 is selected from ISIS NOs: 410544, 410608, 410686, 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, 410692, 410548, 410614, or 410693.

In certain embodiments, a target region is nucleotides 1852-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1852-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 379, 380, 381, 382, 383, 384, 385, or 386. In certain such embodiments, an antisense compound targeted to nucleotides 1852-1879 of SEQ ID NO: 1 is selected from ISIS NOs: 410544, 410608, 410686, 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, 410692, 410548, 410614, 410693, or 405990.

In certain embodiments, a target region is nucleotides 1853-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1853-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 380, 381, 382, 383, 384, 385 or 386. In certain such embodiments, an antisense compound targeted to nucleotides 1853-1879 of SEQ ID NO: 1 is selected from ISIS NOs: 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, 410692, 410548, 410614, 410693, or 405990.

In certain embodiments, a target region is nucleotides 1854-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1854-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 381, 382, 383, 384, 385, or 386. In certain such embodiments, an antisense compound targeted to nucleotides 1854-1879 of SEQ ID NO: 1 is selected from ISIS NOs: 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, 410692, 410548, 410614, 410693, or 405990.

In certain embodiments, a target region is nucleotides 1905-1955 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1905-1955 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ IL NOs: 59, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, or 406. In certain such embodiments, an antisense compound targeted to nucleotides 1905-1955 of SEQ ID NO: 1 is selected from ISIS NOs: 410773, 410549, 410615, 410694, 410550, 410616, 410695, 410551, 410617, 410696, 410552, 410618, 410697, 410553, 410619, 410698, 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, or 410774.

In certain embodiments, a target region is nucleotides 1915-1942 of SEQ IL NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1915-1942 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 388, 389, 390, 391, 392, 393, 394, or 395. In certain such embodiments, an antisense compound targeted to nucleotides 1915-1942 of SEQ ID NO: 1 is selected from ISIS NOs: 410549, 410615, 410694, 410550, 410616, 410695, 410551, 410617, 410696, 410552, 410618, 410697, 410553, 410619, 410698, 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, or 410702.

In certain embodiments, a target region is nucleotides 1916-1943 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1916-1943 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 389, 390, 391, 392, 393, 394, 395, or 396. In certain such embodiments, an antisense compound targeted to nucleotides 1916-1943 of SEQ ID NO: 1 is selected from ISIS NOs: 410550, 410616, 410695, 410551, 410617, 410696, 410552, 410618, 410697, 410553, 410619, 410698, 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, or 410703.

In certain embodiments, a target region is nucleotides 1917-1944 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1917-1944 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 390, 391, 392, 393, 394, 395, 396, or 397. In certain such embodiments, an antisense compound targeted to nucleotides 1917-1944 of SEQ ID NO: 1 is selected from ISIS NOs: 410551, 410617, 410696, 410552, 410618, 410697, 410553, 410619, 410698, 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, or 410704.

In certain embodiments, a target region is nucleotides 1918-1945 of SEQ ID NO:1. In certain embodiments, an antisense compound is targeted to nucleotides 1918-1945 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 391, 392, 393, 394, 395, 396, 397, or 398. In certain such embodiments, an antisense compound targeted to nucleotides 1918-1945 of SEQ ID NO: 1 is selected from ISIS NOs: 410552, 410618, 410697, 410553, 410619, 410698, 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, or 410705.

In certain embodiments, a target region is nucleotides 1919-1946 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1919-1946 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 392, 393, 394, 395, 396, 397, or 399. In certain such embodiments, an antisense compound targeted to nucleotides 1919-1946 of SEQ ID NO: 1 is selected from ISIS NOs: 410553, 410619, 410698, 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, or 410706.

In certain embodiments, a target region is nucleotides 1920-1939 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1920-1939 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59. In certain such embodiments, an antisense compound targeted to nucleotides 1920-1939 of SEQ ID NO: 1 is selected from ISIS NOs: 395185, 399907, or 410699.

In certain embodiments, a target region is nucleotides 1920-1947 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1920-1947 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 393, 394, 395, 396, 397, 398, 399, or 400. In certain such embodiments, an antisense compound targeted to nucleotides 1920-1947 of SEQ ID NO: 1 is selected from ISIS NOs: 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405944, 410627, or 410707.

In certain embodiments, a target region is nucleotides 1921-1948 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1921-1948 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 393, 394, 395, 396, 397, 398, 399, 400, or 401. In certain such embodiments, an antisense compound targeted to nucleotides 1921-1948 of SEQ ID NO: 1 is selected from ISIS NOs: 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405944, 410627, 410707, 410558, 410628, or 410708.

In certain embodiments, a target region is nucleotides 1922-1949 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1922-1949 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 394, 395, 396, 397, 398, 399, 400, 401, or 402. In certain such embodiments, an antisense compound targeted to nucleotides 1922-1949 of SEQ ID NO: 1 is selected from ISIS NOs: 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405944, 410627, 410707, 410558, 410628, 410708, 405995, 410629, or 410709.

In certain embodiments, a target region is nucleotides 1923-1950 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1923-1950 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 395, 396, 397, 398, 399, 400, 401, 402, or 403. In certain such embodiments, an antisense compound targeted to nucleotides 1923-1950 of SEQ ID NO: 1 is selected from ISIS NOs: 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405944, 410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559, 410630, or 410710.

In certain embodiments, a target region is nucleotides 1924-1951 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1924-1951 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 396, 397, 398, 399, 400, 401, 402, 403, or 404. In certain such embodiments, an antisense compound targeted to nucleotides 1924-1951 of SEQ ID NO: 1 is selected from ISIS NOs: 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405994, 410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559, 410630, 410710, 410560, 410631, or 410711.

In certain embodiments, a target region is nucleotides 1925-1952 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1925-1952 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 397, 398, 399, 400, 401, 402, 403, 404, or 405. In certain such embodiments, an antisense compound targeted to nucleotides 1925-1952 of SEQ ID NO: 1 is selected from ISIS NOs: 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405994, 410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559, 410630, 410710, 410560, 410631, 410711, 410561, 410632, or 410712.

In certain embodiments, a target region is nucleotides 1926-1952 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1926-1952 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 398, 399, 400, 401, 402, 403, 404, or 405. In certain such embodiments, an antisense compound targeted to nucleotides 1926-1952 of SEQ ID NO: 1 is selected from ISIS NOs: 405993, 410625, 410705, 410557, 410626, 410706, 405994, 410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559, 410630, 410710, 410560, 410631, 410711, 410561, 410632, or 410712.

In certain embodiments, a target region is nucleotides 1927-1952 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1927-1952 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 399, 400, 401, 402, 403, 404, or 405. In certain such embodiments, an antisense compound targeted to nucleotides 1927-1952 of SEQ ID NO: 1 is selected from ISIS NOs: 410557, 410626, 410706, 405994, 410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559, 410630, 410710, 410560, 410631, 410711, 410561, 410632, or 410712.

In certain embodiments, a target region is nucleotides 1928-1955 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1928-1955 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 400, 401, 402, 403, 404, 405, or 406. In certain such embodiments, an antisense compound targeted to nucleotides 1928-1955 of SEQ ID NO: 1 is selected from ISIS NOs: 405994, 410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559, 410630, 410710, 410560, 410631, 410711, 410561, 410632, 410712, or 410774.

In certain embodiments, a target region is nucleotides 1962-2059 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1962-2059 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 407, 408, 409, or 410. In certain such embodiments, an antisense compound targeted to nucleotides 1962-2059 of SEQ ID NO: 1 is selected from ISIS NOs: 410775, 410776, 410777, or 410778.

In certain embodiments, a target region is nucleotides 2040-2126 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2040-2126 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 410, 411, 412, 413, 414, or 415. In certain such embodiments, an antisense compound targeted to nucleotides 2040-2126 of SEQ ID NO: 1 is selected from ISIS NOs: 410778, 395186, 399908, 410713, 410562, 410633, 410714, 405996, 410634, 410715, 410563, 410635, 410716, 410564, 410636, 410717, 399817, 399973, 410718, or 405997.

In certain embodiments, a target region is nucleotides 2100-2126 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2100-2126 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413, 414, or 415. In certain such embodiments, an antisense compound targeted to nucleotides 2100-2126 of SEQ ID NO: 1 is selected from ISIS NOs: 395186, 399908, 410713, 410562, 410633, 410714, 405996, 410634, 410715, 410563, 410635, 410716, 410564, 410636, 410717, 399817, 399973, 410718 or 405997.

In certain embodiments, a target region is nucleotides 2100-2139 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2100-2139 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413, 414, 415, or 416. In certain such embodiments, an antisense compound targeted to nucleotides 2100-2139 of SEQ ID NO: 1 is selected from ISIS NOs: 395186, 399908, 410713, 410562, 410633, 410714, 405996, 410634, 410715, 410563, 410635, 410716, 410564, 410636, 410717, 399817, 399973, 410718, 405997 or 410779.

In certain embodiments, a target region is nucleotides 2100-2206 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2100-2206 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413, 414, 415, 416, 417, or 418. In certain such embodiments, an antisense compound targeted to nucleotides 2100-2206 of SEQ ID NO: 1 is selected from ISIS NOs: 395186, 399908, 410713, 410562, 410633, 410714, 405996, 410634, 410715, 410563, 410635, 410716, 410564, 410636, 410717, 399817, 399973, 410718, 405997, 405997, 410780, or 410781.

In certain embodiments, a target region is nucleotides 2101-2126 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2101-2126 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 61, 411, 412, 413, 414, or 415. In certain such embodiments, an antisense compound targeted to nucleotides 2101-2126 of SEQ ID NO: 1 is selected from ISIS NOs: 410562, 410633, 410714, 405996, 410634, 410715, 410563, 410635, 410716, 410564, 410636, 410717, 399817, 399973, 410718, or 405997.

In certain embodiments, a target region is nucleotides 2305-2332 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2305-2332 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422, 423, 424, 425, or 426. In certain such embodiments, an antisense compound targeted to nucleotides 2305-2332 of SEQ ID NO: 1 is selected from ISIS NOs: 410565, 410637, 410719, 410566, 410638, 410720, 410567, 410639, 410721, 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, or 410727.

In certain embodiments, a target region is nucleotides 2305-2354 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2305-2354 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, or 430. In certain such embodiments, an antisense compound targeted to nucleotides 2305-2354 of SEQ ID NO: 1 is selected from ISIS NOs: 410565, 410637, 410719, 410566, 410638, 410720, 410567, 410639, 410721, 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410783.

In certain embodiments, a target region is nucleotides 2306-2333 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2306-2333 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 420, 421, 422, 423, 424, 425, 426, or 427. In certain such embodiments, an antisense compound targeted to nucleotides 2306-2333 of SEQ ID NO: 1 is selected from ISIS NOs: 410566, 410638, 410720, 410567, 410639, 410721, 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, 410727, 410572, 410645, or 410728.

In certain embodiments, a target region is nucleotides 2307-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2307-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 421, 422, 423, 424, 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2307-2334 of SEQ ID NO: 1 is selected from ISIS NOs: 410567, 410639, 410721, 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2308-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2308-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 422, 423, 424, 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2308-2334 of SEQ ID NO: 1 is selected from ISIS NOs: 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2309-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2309-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 423, 424, 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2309-2334 of SEQ ID NO: 1 is selected from ISIS NOs: 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2310-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2310-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 424, 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2310-2334 of SEQ ID NO: 1 is selected from ISIS NOs: 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2410-2434 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2410-2434 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 63 or 154. In certain such embodiments, an antisense compound targeted to nucleotides 2410-2434 of SEQ ID NO: 1 is selected from ISIS NOs: 395188, 399910, 399818, or 399974.

In certain embodiments, a target region is nucleotides 2504-2528 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2504-2528 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 64 or 65. In certain such embodiments, an antisense compound targeted to nucleotides 2504-2528 of SEQ ID NO: 1 is selected from ISIS NOs: 395189, 399911, 399819, or 399975.

In certain embodiments, a target region is nucleotides 2509-2528 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2509-2528 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 65. In certain such embodiments, an antisense compound targeted to nucleotides 2509-2528 of SEQ ID NO: 1 is selected from ISIS NOs: 399819 or 399975.

In certain embodiments, a target region is nucleotides 2582-2625 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2582-2625 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 66, 67 or 122. In certain such embodiments, an antisense compound targeted to nucleotides 2582-2625 of SEQ ID NO: 1 is selected from ISIS NOs: 399820, 399976, 395190, 399912, 395191, or 399913.

In certain embodiments, a target region is nucleotides 2606-2668 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2606-2668 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 67 or 153. In certain such embodiments, an antisense compound targeted to nucleotides 2606-2668 of SEQ ID NO: 1 is selected from ISIS NOs: 395191, 399913, 395192, or 399914.

In certain embodiments, a target region is nucleotides 2828-2855 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2828-2855 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 69, 431, 432, 433, 434, 435, 436, 437 or 438. In certain such embodiments, an antisense compound targeted to nucleotides 2828-2855 of SEQ ID NO: 1 is selected from ISIS NOs: 405893, 405894, 405895, 405896, 395194, 399916, 405897, 405898, 405899, or 405900.

In certain embodiments, a target region is nucleotides 2832-2851 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2832-2851 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 69. In certain such embodiments, an antisense compound targeted to nucleotides 2832-2851 of SEQ ID NO: 1 is selected from ISIS NOs: 395194, or 399916.

In certain embodiments, a target region is nucleotides 2900-2927 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2900-2927 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441, 442, 443 or 444. In certain such embodiments, an antisense compound targeted to nucleotides 2900-2927 of SEQ ID NO: 1 is selected from ISIS NOs: 395195, 399917, 405901, 405902, 405903, 405904, 399821, 399977, 405905, or 405906.

In certain embodiments, a target region is nucleotides 2900-2929 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2900-2929 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441, 442, 443, 444, 446 or 446. In certain such embodiments, an antisense compound targeted to nucleotides 2900-2929 of SEQ ID NO: 1 is selected from ISIS NOs: 395195, 399917, 405901, 405902, 405903, 405904, 399821, 399977, 405905, 405906, 405907, or 405908.

In certain embodiments, a target region is nucleotides 2902-2927 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2902-2927 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 71, 439, 440, 441, 442, 443 or 444. In certain such embodiments, an antisense compound targeted to nucleotides 2902-2927 of SEQ ID NO: 1 is selected from ISIS NOs: 405901, 405902, 405903, 405904, 399821, 399977, 405905, or 405906.

In certain embodiments, a target region is nucleotides 2983-3007 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2983-3007 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 72 or 73. In certain such embodiments, an antisense compound targeted to nucleotides 2983-3007 of SEQ ID NO: 1 is selected from ISIS NOs: 395196, 399918, 399822, or 399978.

In certain embodiments, a target region is nucleotides 2983-3013 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2983-3013 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 72, 73 or 135. In certain such embodiments, an antisense compound targeted to nucleotides 2983-3013 of SEQ ID NO: 1 is selected from ISIS NOs: 395196, 399918, 399822, 399978, 399823, or 399979.

In certain embodiments, a target region is nucleotides 3227-3252 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 3227-3252 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 74 or 112. In certain such embodiments, an antisense compound targeted to nucleotides 3227-3252 of SEQ ID NO: 1 is selected from ISIS NOs: 395197, 399919, 399824, or 399980.

In certain embodiments, a target region is nucleotides 3227-3456 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 3227-3456 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 74, 75 or 112. In certain such embodiments, an antisense compound targeted to nucleotides 3227-3456 of SEQ ID NO: 1 is selected from ISIS NOs: 395197, 399919, 399824, 399980, 395198, or 399920.

In certain embodiments, a target region is nucleotides 3472-3496 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 3472-3496 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 76 or 77. In certain such embodiments, an antisense compound targeted to nucleotides 3472-3496 of SEQ ID NO: 1 is selected from ISIS NOs: 395199, 399921, 399825 or 399981.

In certain embodiments, a target region is nucleotides 3543-3569 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 3543-3569 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 78 or 99. In certain such embodiments, an antisense compound targeted to nucleotides 3543-3569 of SEQ ID NO: 1 is selected from ISIS NOs: 395200, 399922, 399826 or 399982.

In certain embodiments, antisense compound target a PCSK9 nucleic acid having the sequence of nucleotides 25475000 to 25504000 of GENBANK® Accession No. NT_(—)032977.8, first deposited with GENBANK® on Feb. 26, 2006, and incorporated herein as SEQ ID NO: 2. In certain such embodiments, an antisense oligonucleotide is targeted to SEQ ID NO: 2. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 2 is at least 90% complementary to SEQ ID NO: 2. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 2 is at least 95% complementary to SEQ ID NO: 2. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 1 is 100% complementary to SEQ ID NO: 1. In certain such embodiments, an antisense oligonucleotide comprises a nucleotide sequence selected from a nucleotide sequence set forth in Table 7.

TABLE 7 Antisense sequences targeted to NT_032977.8 (SEQ ID NO: 2) 5′ Start 3′ Start Site to Site to SEQ ID SEQ ID SEQ ID NO NO: 2 NO: 2 Sequence (5′ to 3′) 4 2274 2293 GCGCGGAATCCTGGCTGGGA 5 2381 2400 GAGGAGACCTAGAGGCCGTG 159 2433 2452 GCCTGGAGCTGACGGTGCCC 160 2437 2456 GACCGCCTGGAGCTGACGGT 6 2439 2458 AGGACCGCCTGGAGCTGACG 162 2545 2564 AAGGCTAGCACCAGCTCCTC 163 2546 2565 CAAGGCTAGCACCAGCTCCT 164 2547 2566 GCAAGGCTAGCACCAGCTCC 165 2548 2567 CGCAAGGCTAGCACCAGCTC 7 2549 2568 ACGCAAGGCTAGCACCAGCT 166 2550 2569 AACGCAAGGCTAGCACCAGC 167 2551 2570 GAACGCAAGGCTAGCACCAG 168 2552 2571 GGAACGCAAGGCTAGCACCA 169 2553 2572 CGGAACGCAAGGCTAGCACC 8 2556 2575 CCTCGGAACGCAAGGCTAGC 170 2560 2579 TCCTCCTCGGAACGCAAGGC 171 2585 2604 GTGCTCGGGTGCTTCGGCCA 172 2605 2624 TGGAAGGTGGCTGTGGTTCC 9 2619 2638 CCTTGGCGCAGCGGTGGAAG 107 3056 3075 CCCACTATAATGGCAAGCCC 80 4306 4325 AACCCAGTTCTAATGCACCT 106 5140 5159 CCAGTCAGAGTAGAACAGAG 102 5590 5609 ATGTGCAGAGATCAATCACA 121 5599 5618 GGAGCCTACATGTGCAGAGA 94 5667 5686 AGCATGGCACCAGCATCTGC 176 6444 6463 CGTAGGTGCCAGGCAACCTC 177 6482 6501 TGACTGCGAGAGGTGGGTCT 178 6492 6511 CAGTGCGCTCTGACTGCGAG 179 6494 6513 GGCAGTGCGCTCTGACTGCG 180 6496 6515 CGGGCAGTGCGCTCTGACTG 10 6498 6517 GGCGGGCAGTGCGCTCTGAC 181 6499 6518 CGGCGGGCAGTGCGCTCTGA 182 6528 6547 ATCCCCGGCGGGCAGCCTGG 183 6532 6551 AGGTATCCCCGGCGGGCAGC 184 6534 6553 TGAGGTATCCCCGGCGGGCA 185 6535 6554 GTGAGGTATCCCCGGCGGGC 186 6536 6555 GGTGAGGTATCCCCGGCGGG 11 6537 6556 TGGTGAGGTATCCCCGGCGG 187 6538 6557 TTGGTGAGGTATCCCCGGCG 188 6539 6558 CTTGGTGAGGTATCCCCGGC 189 6540 6559 TCTTGGTGAGGTATCCCCGG 190 6541 6560 ATCTTGGTGAGGTATCCCCG 191 6542 6561 GATCTTGGTGAGGTATCCCC 12 6543 6562 GGATCTTGGTGAGGTATCCC 192 6544 6563 AGGATCTTGGTGAGGTATCC 193 6546 6565 GCAGGATCTTGGTGAGGTAT 194 6548 6567 ATGCAGGATCTTGGTGAGGT 195 6550 6569 ACATGCAGGATCTTGGTGAG 13 6552 6571 AGACATGCAGGATCTTGGTG 196 6554 6573 GAAGACATGCAGGATCTTGG 14 6557 6576 ATGGAAGACATGCAGGATCT 197 6565 6584 AGAAGGCCATGGAAGACATG 198 6575 6594 GAAGCCAGGAAGAAGGCCAT 15 6583 6602 TTCACCAGGAAGCCAGGAAG 199 6585 6604 TCTTCACCAGGAAGCCAGGA 16 6588 6607 TCATCTTCACCAGGAAGCCA 200 6590 6609 ACTCATCTTCACCAGGAAGC 201 6592 6611 CCACTCATCTTCACCAGGAA 202 6594 6613 CGCCACTCATCTTCACCAGG 203 6596 6615 GTCGCCACTCATCTTCACCA 204 6598 6617 AGGTCGCCACTCATCTTCAC 205 6600 6619 GCAGGTCGCCACTCATCTTC 206 6602 6621 CAGCAGGTCGCCACTCATCT 207 6604 6623 TCCAGCAGGTCGCCACTCAT 108 6652 6671 CCCAGCCCTATCAGGAAGTG 144 7099 7118 TGACATCCAGGAGGGAGGAG 91 7556 7575 AGACTGATGGAAGGCATTGA 131 7565 7584 GTGTTGAGCAGACTGATGGA 145 8836 8855 TGACATCTTGTCTGGGAGCC 90 8948 8967 AGACTAGGAGCCTGAGTTTT 125 9099 9118 GGCCTGCAGAAGCCAGAGAG 17 9130 9149 CCTCGATGTAGTCGACATGG 209 9149 9168 GCAAAGACAGAGGAGTCCTC 210 9207 9226 TTCATCCGCCCGGTACCGTG 211 9209 9228 TATTCATCCGCCCGGTACCG 18 9210 9229 GTATTCATCCGCCCGGTACC 212 9212 9231 TGGTATTCATCCGCCCGGTA 213 9214 9233 GCTGGTATTCATCCGCCCGG 214 9216 9235 GGGCTGGTATTCATCCGCCC 148 10252 10271 TGGCAGCAACTCAGACATAT 127 10633 10652 GGTGGTAATTTGTCACAGCA 84 11308 11327 AAGGTCACACAGTTAAGAGT 79 11472 11491 AAATGCAGGGCTAAAATCAC 88 12715 12734 ACTGGATACATTGGCAGACA 111 12928 12947 CTAGAGGAACCACTAGATAT 85 13681 13700 ACAAATTCCCAGACTCAGCA 100 13746 13765 ATCTCAGGACAGGTGAGCAA 116 13760 13779 GAGTAGAGATTCTCATCTCA 129 13816 13835 GTGCCATCTGAACAGCACCT 117 13828 13847 GAGTCTTCTGAAGTGCCATC 81 13903 13922 AAGCAGGGCCTCAGGTGGAA 110 13926 13945 CCTGGAACCCCTGCAGCCAG 152 13977 13996 TTCAGGCAGGTTGCTGCTAG 83 13986 14005 AAGGAAGACTTCAGGCAGGT 140 13998 14017 TCAGCCAGGCCAAAGGAAGA 137 14112 14131 TAGGGAGAGCTCACAGATGC 136 14122 14141 TAGGAGAAAGTAGGGAGAGC 132 14179 14198 TAAAAGCTGCAAGAGACTCA 139 14267 14286 TCAGAGAAAACAGTCACCGA 92 14397 14416 AGAGACAGGAAGCTGCAGCT 142 14404 14423 TCATTTTAGAGACAGGAAGC 113 14441 14460 GAATAACAGTGATGTCTGGC 138 14494 14513 TCACAGCTCACCGAGTCTGC 98 14524 14543 AGTGTAAAATAAAGCCCCTA 96 14601 14620 AGGACCCAAGTCATCCTGCT 124 14631 14650 GGCCATCAGCTGGCAATGCT 82 14670 14689 AAGGAAAGGGAGGCCTAGAG 133 14675 14694 TAGACAAGGAAAGGGAGGCC 103 14681 14700 ATTTCATAGACAAGGAAAGG 155 14801 14820 CTTATAGTTAACACACAGAA 156 14809 14828 AAGTCAACCTTATAGTTAAC 215 14877 14896 ATACACCTCCACCAGGCTGC 216 14888 14907 GTGTCTAGGAGATACACCTC 19 14891 14910 CTGGTGTCTAGGAGATACAC 217 14893 14912 TGCTGGTGTCTAGGAGATAC 20 14896 14915 GTATGCTGGTGTCTAGGAGA 21 14916 14935 GATTTCCCGGTGGTCACTCT 218 14918 14937 TCGATTTCCCGGTGGTCACT 219 14919 14938 CTCGATTTCCCGGTGGTCAC 220 14920 14939 CCTCGATTTCCCGGTGGTCA 221 14921 14940 CCCTCGATTTCCCGGTGGTC 22 14922 14941 GCCCTCGATTTCCCGGTGGT 222 14923 14942 TGCCCTCGATTTCCCGGTGG 223 14924 14943 CTGCCCTCGATTTCCCGGTG 224 14925 14944 CCTGCCCTCGATTTCCCGGT 225 14926 14945 CCCTGCCCTCGATTTCCCGG 226 14930 14949 ATGACCCTGCCCTCGATTTC 227 14932 14951 CCATGACCCTGCCCTCGATT 228 14934 14953 GACCATGACCCTGCCCTCGA 23 14936 14955 GTGACCATGACCCTGCCCTC 229 14938 14957 CGGTGACCATGACCCTGCCC 230 14940 14959 GTCGGTGACCATGACCCTGC 231 14942 14961 AAGTCGGTGACCATGACCCT 232 14944 14963 CGAAGTCGGTGACCATGACC 24 14946 14965 CTCGAAGTCGGTGACCATGA 233 14954 14973 GGCACATTCTCGAAGTCGGT 25 14979 14998 GTGGAAGCGGGTCCCGTCCT 235 15254 15273 GGTGGGTGCCATGACTGTCA 236 15257 15276 CCAGGTGGGTGCCATGACTG 237 15261 15280 CCTGCCAGGTGGGTGCCATG 26 15264 15283 ACCCCTGCCAGGTGGGTGCC 238 15266 15285 CCACCCCTGCCAGGTGGGTG 27 15269 15288 TGACCACCCCTGCCAGGTGG 239 15271 15290 GCTGACCACCCCTGCCAGGT 240 15279 15298 TCCCGGCCGCTGACCACCCC 241 15283 15302 GGCATCCCGGCCGCTGACCA 242 15286 15305 GCCGGCATCCCGGCCGCTGA 243 15291 15310 GCCACGCCGGCATCCCGGCC 244 15292 15311 GGCCACGCCGGCATCCCGGC 245 15293 15312 TGGCCACGCCGGCATCCCGG 246 15294 15313 TTGGCCACGCCGGCATCCCG 247 15295 15314 CTTGGCCACGCCGGCATCCC 248 15296 15315 CCTTGGCCACGCCGGCATCC 249 15297 15316 CCCTTGGCCACGCCGGCATC 28 15298 15317 ACCCTTGGCCACGCCGGCAT 447 15298 15317 ACCCTTGGTCACGCCGGCAT 250 15299 15318 CACCCTTGGCCACGCCGGCA 251 15300 15319 GCACCCTTGGCCACGCCGGC 252 15301 15320 GGCACCCTTGGCCACGCCGG 253 15302 15321 TGGCACCCTTGGCCACGCCG 254 15303 15322 CTGGCACCCTTGGCCACGCC 255 15304 15323 GCTGGCACCCTTGGCCACGC 256 15309 15328 CGCATGCTGGCACCCTTGGC 257 15330 15349 CAGTTGAGCACGCGCAGGCT 258 15332 15351 GGCAGTTGAGCACGCGCAGG 29 15334 15353 TTGGCAGTTGAGCACGCGCA 259 15336 15355 CCTTGGCAGTTGAGCACGCG 30 15339 15358 TTCCCTTGGCAGTTGAGCAC 260 15341 15360 CCTTCCCTGGCAGTTGAGC 261 15345 15364 GTGCCCTTCCCTTGGCAGTT 262 15347 15366 CCGTGCCCTTCCCTTGGCAG 263 15358 15377 GGTGCCGCTAACCGTGCCCT 86 15471 15490 ACAGCATTCTTGGTTAGGAG 97 16134 16153 AGTCAAGCTGCTGCCCAGAG 120 16668 16687 GCTAGTTATTAAGCACCTGC 150 17267 17286 TGTGAGCTCTGGCCCAGTGG 115 18377 18396 GAGTAAGGCAGGTTACTCTC 134 18408 18427 TAGATGTGACTAACATTTAA 157 18561 18580 AGGAACAAAGCCAAGGTCAC 266 18591 18610 TGGCTTTTCCGAATAAACTC 32 18593 18612 GCTGGCTTTTCCGAATAAAC 267 18595 18614 CAGCTGGCTTTTCCGAATAA 268 18597 18616 ACCAGCTGGCTTTTCCGAAT 269 18599 18618 GGACCAGCTGGCTTTTCCGA 270 18603 18622 GGCTGGACCAGCTGGCTTTT 271 18614 18633 GTGGCCCCACAGGCTGGACC 272 18627 18646 AGCAGCACCACCAGTGGCCC 273 18649 18668 GCTGTACCCACCCGCCAGGG 274 18695 18714 CGACCCCAGCCCTCGCCAGG 33 18705 18724 GTGACCAGCACGACCCCAGC 275 18707 18726 CGGTGACCAGCACGACCCCA 276 18709 18728 AGCGGTGACCAGCACGACCC 277 18711 18730 GCAGCGGTGACCAGCACGAC 278 18713 18732 CGGCAGCGGTGACCAGCACG 279 18714 18733 CCGGCAGCGGTGACCAGCAC 280 18717 18736 TTGCCGGCAGCGGTGACCAG 281 18719 18738 AGTTGCCGGCAGCGGTGACC 282 18721 18740 GAAGTTGCCGGCAGCGGTGA 283 18723 18742 CGGAAGTTGCCGGCAGCGGT 284 18725 18744 CCCGGAAGTTGCCGGCAGCG 285 18727 18746 GTCCCGGAAGTTGCCGGCAG 105 19203 19222 CACATTAGCCTTGCTCAAGT 151 19913 19932 TGTGATGACCTGGAAAGGTG 288 19931 19950 GGCATTGGTGGCCCCAACTG 149 19933 19952 TGGGCATTGGTGGCCCCAAC 289 19941 19960 GCTGGTCTTGGGCATTGGTG 290 19954 19973 CCCAGGGTCACCGGCTGGTC 291 19956 19975 TCCCCAGGGTCACCGGCTGG 292 19958 19977 AGTCCCCAGGGTCACCGGCT 293 19960 19979 AAAGTCCCCAGGGTCACCGG 34 19962 19981 CCAAAGTCCCCAGGGTCACC 294 19964 19983 CCCCAAAGTCCCCAGGGTCA 128 19966 19985 GTCCCCAAAGTCCCCAGGGT 295 19969 19988 TTGGTCCCCAAAGTCCCCAG 35 19971 19990 AGTTGGTCCCCAAAGTCCCC 296 19973 19992 AAAGTTGGTCCCCAAAGTCC 36 19976 19995 GCCAAAGTTGGTCCCCAAAG 297 19978 19997 CGGCCAAAGTTGGTCCCCAA 298 19980 19999 AGCGGCCAAAGTTGGTCCCC 299 19982 20001 ACAGCGGCCAAAGTTGGTCC 300 19984 20003 ACACAGCGGCCAAAGTTGGT 301 19986 20005 CCACACAGCGGCCAAAGTTG 37 19988 20007 GTCCACACAGCGGCCAAAGT 302 19990 20009 AGGTCCACACAGCGGCCAAA 303 19992 20011 AGAGGTCCACACAGCGGCCA 304 19994 20013 AAAGAGGTCCACACAGCGGC 38 19997 20016 GGCAAAGAGGTCCACACAGC 305 20016 20035 CAATGATGTCCTCCCCTGGG 306 20023 20042 GAGGCACCAATGATGTCCTC 39 20025 20044 TGGAGGCACCAATGATGTCC 307 20027 20046 GCTGGAGGCACCAATGATGT 308 20029 20048 TCGCTGGAGGCACCAATGAT 309 20031 20050 AGTCGCTGGAGGCACCAATG 310 20033 20052 GCAGTCGCTGGAGGCACCAA 40 20036 20055 GCTGCAGTCGCTGGAGGCAC 311 20038 20057 GTGCTGCAGTCGCTGGAGGC 312 20040 20059 AGGTGCTGCAGTCGCTGGAG 313 20042 20061 GCAGGTGCTGCAGTCGCTGG 314 20043 20062 AGCAGGTGCTGCAGTCGCTG 315 20045 20064 AAAGCAGGTGCTGCAGTCGC 41 20047 20066 ACAAAGCAGGTGCTGCAGTC 316 20049 20068 ACACAAAGCAGGTGCTGCAG 317 20051 20070 TGACACAAAGCAGGTGCTGC 318 20061 20080 TCCCACTCTGTGACACAAAG 158 20100 20119 GTGGTGACTTACCAGCCACG 109 20188 20207 CCCCTGCACAGAGCCTGGCA 141 20624 20643 TCATGGCTGCAATGCCTGGT 320 20629 20648 CAGCATCATGGCTGCAATGC 321 20631 20650 GACAGCATCATGGCTGCAAT 322 20633 20652 CAGACAGCATCATGGCTGCA 43 20635 20654 GGCAGACAGCATCATGGCTG 323 20637 20656 TCGGCAGACAGCATCATGGC 324 20639 20658 GCTCGGCAGACAGCATCATG 325 20641 20660 CGGCTCGGCAGACAGCATCA 326 20643 20662 TCCGGCTCGGCAGACAGCAT 327 20657 20676 CGGCCAGGGTGAGCTCCGGC 328 20670 20689 CTCTGCCTCAACTCGGCCAG 329 20672 20691 GTCTCTGCCTCAACTCGGCC 330 20674 20693 CAGTCTCTGCCTCAACTCGG 331 20676 20695 ATCAGTCTCTGCCTCAACTC 332 20678 20697 GGATCAGTCTCTGCCTCAAC 333 20680 20699 GTGGATCAGTCTCTGCCTCA 334 20682 20701 AAGTGGATCAGTCTCTGCCT 44 20683 20702 GAAGTGGATCAGTCTCTGCC 335 20685 20704 GAGAAGTGGATCAGTCTCTG 336 20687 20706 CAGAGAAGTGGATCAGTCTC 337 20689 20708 GGCAGAGAAGTGGATCAGTC 45 20691 20710 TTGGCAGAGAAGTGGATCAG 338 20693 20712 CTTTGGCAGAGAAGTGGATC 46 20696 20715 CATCTTTGGCAGAGAAGTGG 339 20698 20717 GACATCTTTGGCAGAGAAGT 340 20700 20719 ATGACATCTTTGGCAGAGAA 47 20702 20721 TGATGACATCTTTGGCAGAG 341 20704 20723 ATTGATGACATCTTTGGCAG 342 20706 20725 TCATTGATGACATCTTTGGC 48 20709 20728 GCCTCATTGATGACATCTTT 343 20711 20730 AGGCCTCATTGATGACATCT 344 20713 20732 CCAGGCCTCATTGATGACAT 345 20715 20734 AACCAGGCCTCATTGATGAC 346 20717 20736 GGAACCAGGCCTCATTGATG 347 20718 20737 GGGAACCAGGCCTCATTGAT 348 20719 20738 AGGGAACCAGGCCTCATTGA 349 20720 20739 CAGGGAACCAGGCCTCATTG 49 20721 20740 TCAGGGAACCAGGCCTCATT 350 20722 20741 CTCAGGGAACCAGGCCTCAT 351 20723 20742 CCTCAGGGAACCAGGCCTCA 352 20724 20743 TCCTCAGGGAACCAGGCCTC 353 20725 20744 GTCCTCAGGGAACCAGGCCT 50 20726 20745 GGTCCTCAGGGAACCAGGCC 354 20727 20746 TGGTCCTCAGGGAACCAGGC 355 20728 20747 CTGGTCCTCAGGGAACCAGG 356 20729 20748 GCTGGTCCTCAGGGAACCAG 357 20730 20749 CGCTGGTCCTCAGGGAACCA 87 20733 20752 ACCCGCTGGTCCTCAGGGAA 358 20735 20754 GTACCCGCTGGTCCTCAGGG 359 20737 20756 CAGTACCCGCTGGTCCTCAG 51 20740 20759 GGTCAGTACCCGCTGGTCCT 119 20762 20781 GCAGGGCGGCCACCAGGTTG 360 20785 20804 ACCTGCCCCATGGGTGCTGG 93 20995 21014 AGAGAGGAGGGCTTAAAGAA 95 21082 21101 AGCTGCCAACCTGCAAAAAG 361 21088 21107 CAAAACAGCTGCCAACCTGC 53 21091 21110 CTGCAAAACAGCTGCCAACC 362 21093 21112 TCCTGCAAAACAGCTGCCAA 363 21095 21114 AGTCCTGCAAAACAGCTGCC 364 21106 21125 GCTGACCATACAGTCCTGCA 365 21118 21137 GGCCCCGAGTGTGCTGACCA 54 21121 21140 GTAGGCCCCGAGTGTGCTGA 366 21123 21142 GTGTAGGCCCCGAGTGTGCT 367 21125 21144 CCGTGTAGGCCCCGAGTGTG 368 21127 21146 ATCCGTGTAGGCCCCGAGTG 369 21129 21148 CCATCCGTGTAGGCCCCGAG 370 21131 21150 GGCCATCCGTGTAGGCCCCG 371 21133 21152 GTGGCCATCCGTGTAGGCCC 372 21181 21200 CTGGAGCAGCTCAGCAGCTC 460 21181 21194 CAGCTCAGCAGCTC 373 21183 21202 AACTGGAGCAGCTCAGCAGC 55 21186 21205 AGAAACTGGAGCAGCTCAGC 374 21188 21207 GGAGAAACTGGAGCAGCTCA 375 21190 21209 CTGGAGAAACTGGAGCAGCT 56 21192 21211 TCCTGGAGAAACTGGAGCAG 143 21481 21500 TGAAAATCCATCCAGCACTG 89 21589 21608 AGAACCATGGAGCACCTGAG 448 21692 21711 CTGCCCTTCCACCAAAATGC 449 21693 21712 ACTGCCCTTCCACCAAAATG 450 21694 21713 CACTGCCCTTCCACCAAAAT 451 21695 21714 GCACTGCCCTTCCACCAAAA 123 21696 21715 GGCACTGCCCTTCCACCAAA 452 21697 21716 GGGCACTGCCCTTCCACCAA 453 21698 21717 TGGGCACTGCCCTTCCACCA 454 21699 21718 CTGGGCACTGCCCTTCCACC 455 21700 21719 GCTGGGCACTGCCCTTCCAC 57 22096 22115 CGTTGTGGGCCCGGCAGACC 376 22133 22152 CACCTGGCAATGGCGTAGAC 377 22134 22153 GCACCTGGCAATGGCGTAGA 378 22135 22154 AGCACCTGGCAATGGCGTAG 379 22136 22155 CAGCACCTGGCAATGGCGTA 380 22137 22156 GCAGCACCTGGCAATGGCGT 381 22138 22157 GGCAGCACCTGGCAATGGCG 382 22139 22158 AGGCAGCACCTGGCAATGGC 383 22140 22159 CAGGCAGCACCTGGCAATGG 384 22141 22160 GCAGGCAGCACCTGGCAATG 58 22142 22161 AGCAGGCAGCACCTGGCAAT 385 22143 22162 TAGCAGGCAGCACCTGGCAA 386 22144 22163 GTAGCAGGCAGCACCTGGCA 387 22189 22208 TGGCCTCAGCTGGTGGAGCT 388 22199 22218 GTCCCCATGCTGGCCTCAGC 389 22200 22219 GGTCCCCATGCTGGCCTCAG 390 22201 22220 GGGTCCCCATGCTGGCCTCA 391 22202 22221 CGGGTCCCCATGCTGGCCTC 392 22203 22222 ACGGGTCCCCATGCTGGCCT 59 22204 22223 CACGGGTCCCCATGCTGGCC 393 22205 22224 ACACGGGTCCCCATGCTGGC 394 22266 22225 GACACGGGTCCCCATGCTGG 395 22207 22226 GGACACGGGTCCCCATGCTG 396 22208 22227 TGGACACGGGTCCCCATGCT 397 22209 22228 GTGGACACGGGTCCCCATGC 398 22210 22229 AGTGGACACGGGTCCCCATG 399 22211 22230 CAGTGGACACGGGTCCCCAT 400 22212 22231 GCAGTGGACACGGGTCCCCA 401 22213 22232 GGCAGTGGACACGGGTCCCC 402 22214 22233 TGGCAGTGGACACGGGTCCC 403 22215 22234 GTGGCAGTGGACACGGGTCC 404 22216 22235 GGTGGCAGTGGACACGGGTC 405 22217 22236 TGGTGGCAGTGGACACGGGT 406 22220 22239 TGTTGGTGGCAGTGGACACG 126 22292 22311 GGTGCATAAGGAGAAAGAGA 408 23985 24004 GTGCCAAGGTCCTCCACCTC 409 24005 24024 TCAGCACAGGCGGCTTGTGG 410 24035 24054 CCACGCACTGGTTGGGCTGA 60 24095 24114 CTTTGCATTCCAGACCTGGG 411 24096 24115 ACTTTGCATTCCAGACCTGG 412 24097 24116 GACTTTGCATTCCAGACCTG 413 24098 24117 TGACTTTGCATTCCAGACCT 414 24099 24118 TTGACTTTGCATTCCAGACC 61 24100 24119 CTTGACTTTGCATTCCAGAC 415 24102 24121 TCCTTGACTTTGCATTCCAG 416 24115 24134 CGGGATTCCATGCTCCTTGA 147 24858 24877 TGAGTTCATTTAAGAGTGGA 118 24907 24926 GCACCATCCAGACCAGAATC 114 25413 25432 GAGAGGTTCAGATCCAGGCC 418 25994 26013 GGAGGGCACTGCAGCCAGTC 419 26112 26131 CAGATGGCAACGGCTGTCAC 420 26113 26132 GCAGATGGCAACGGCTGTCA 421 26114 26133 AGCAGATGGCAACGGCTGTC 422 26115 26134 CAGCAGATGGCAACGGCTGT 423 26116 26135 GCAGCAGATGGCAACGGCTG 62 26117 26136 GGCAGCAGATGGCAACGGCT 424 26118 26137 CGGCAGCAGATGGCAACGGC 425 26119 26138 CCGGCAGCAGATGGCAACGG 426 26120 26139 TCCGGCAGCAGATGGCAACG 427 26121 26140 CTCCGGCAGCAGATGGCAAC 428 26122 26141 GCTCCGGCAGCAGATGGCAA 429 26132 26151 CCAGGTGCCGGCTCCGGCAG 430 26142 26161 GAGGCCTGCGCCAGGTGCCG 154 26217 26236 TTTTAAAGCTCAGCCCCAGC 63 26222 26241 AACCATTTTAAAGCTCAGCC 64 26311 26330 TCAAGGGCCAGGCCAGCAGC 65 26316 26335 CCCACTCAAGGGCCAGGCCA 122 26389 26408 GGAGGGAGCTTCCTGGCACC 66 26404 26423 ATGCCCCACAGTGAGGGAGG 67 26413 26432 AATGGTGAAATGCCCCACAG 153 26456 26475 TTGGGAGCAGCTGGCAGCAC 68 26557 26576 CATGGGAAGAATCCTGCCTC 431 26635 26654 ATGAGGGCCATCAGCACCTT 432 26636 26655 GATGAGGGCCATCAGCACCT 433 26637 26656 AGATGAGGGCCATCAGCACC 434 26638 26657 GAGATGAGGGCCATCAGCAC 69 26639 26658 GGAGATGAGGGCCATCAGCA 435 26640 26659 TGGAGATGAGGGCCATCAGC 436 26641 26660 CTGGAGATGAGGGCCATCAG 437 26642 26661 GCTGGAGATGAGGGCCATCA 438 26643 26662 AGCTGGAGATGAGGGCCATC 461 26684 26697 TTAATCAGGGAGCC 70 26707 26726 TAGATGCCATCCAGAAAGCT 439 26709 26728 GCTAGATGCCATCCAGAAAG 440 26710 26729 GGCTAGATGCCATCCAGAAA 441 26711 26730 TGGCTAGATGCCATCCAGAA 442 26712 26731 CTGGCTAGATGCCATCCAGA 71 26713 26732 TCTGGCTAGATGCCATCCAG 443 26714 26733 CTCTGGCTAGATGCCATCCA 444 26715 26734 CCTCTGGCTAGATGCCATCC 445 26716 26735 GCCTCTGGCTAGATGCCATC 446 26717 26736 AGCCTCTGGCTAGATGCCAT 72 26790 26809 GGCATAGAGCAGAGTAAAGG 73 26795 26814 AGCCTGGCATAGAGCAGAGT 135 26801 26820 TAGCACAGCCTGGCATAGAG 112 27034 27053 GAAGAGGCTTGGCTTCAGAG 74 27040 27059 AAGTAAGAAGAGGCTTGGCT 75 27244 27263 GCTCAAGGAGGGACAGTTGT 76 27279 27298 AAAGATAAATGTCTGCTTGC 77 27284 27303 ACCCAAAAGATAAATGTCTG 78 27350 27369 TCTTCAAGTTACAAAAGCAA 99 27357 27376 ATAAATATCTTCAAGTTACA

In certain embodiments, gapmer antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gapmer antisense compounds are targeted to SEQ ID NO: 2. In certain such embodiments, the nucleotide sequences illustrated in Table 7 have a 5-10-5 gapmer motif. Table 8 illustrates gapmer antisense compounds targeted to SEQ ID NO: 2, having a 5-10-5 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 8 Gapmer antisense compounds having a 5-10-5 motif targeted to SEQ ID NO: 2 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ ID Isis No Motif NO NO: 2 NO: 2 Mismatches Sequence (5′-3′) 395149 5-10-5 4 2274 2293 0 GCGCGGAATCCTGGCTGGGA 395150 5-10-5 5 2381 2400 0 GAGGAGACCTAGAGGCCGTG 395151 5-10-5 6 2439 2458 0 AGGACCGCCTGGAGCTGACG 395152 5-10-5 7 2549 2568 0 ACGCAAGGCTAGCACCAGCT 399793 5-10-5 8 2556 2575 0 CCTCGGAACGCAAGGCTAGC 395153 5-10-5 9 2619 2638 0 CCTTGGCGCAGCGGTGGAAG 399837 5-10-5 107 3056 3075 0 CCCACTATAATGGCAAGCCC 399838 5-10-5 80 4306 4325 0 AACCCAGTTCTAATGCACCT 399839 5-10-5 106 5140 5159 0 CCAGTCAGAGTAGAACAGAG 395221 5-10-5 102 5590 5609 0 ATGTGCAGAGATCAATCACA 399840 5-10-5 121 5599 5618 0 GGAGCCTACATGTGCAGAGA 399841 5-10-5 94 5667 5686 0 AGCATGGCACCAGCATCTGC 395154 5-10-5 10 6498 6517 0 GGCGGGCAGTGCGCTCTGAC 395155 5-10-5 11 6537 6556 0 TGGTGAGGTATCCCCGGCGG 399794 5-10-5 12 6543 6562 0 GGATCTTGGTGAGGTATCCC 399795 5-10-5 13 6552 6571 0 AGACATGCAGGATCTTGGTG 395156 5-10-5 14 6557 6576 0 ATGGAAGACATGCAGGATCT 395157 5-10-5 15 6583 6602 0 TTCACCAGGAAGCCAGGAAG 399796 5-10-5 16 6588 6607 0 TCATCTTCACCAGGAAGCCA 399842 5-10-5 108 6652 6671 0 CCCAGCCCTATCAGGAAGTG 399843 5-10-5 144 7099 7118 0 TGACATCCAGGAGGGAGGAG 399844 5-10-5 91 7556 7575 0 AGACTGATGGAAGGCATTGA 399845 5-10-5 131 7565 7584 0 GTGTTGAGCAGACTGATGGA 399846 5-10-5 145 8836 8855 0 TGACATCTTGTCTGGGAGCC 399847 5-10-5 90 8948 8967 0 AGACTAGGAGCCTGAGTTTT 399848 5-10-5 125 9099 9118 0 GGCCTGCAGAAGCCAGAGAG 395158 5-10-5 17 9130 9149 0 CCTCGATGTAGTCGACATGG 395159 5-10-5 18 9210 9229 0 GTATTCATCCGCCCGGTACC 399849 5-10-5 148 10252 10271 0 TGGCAGCAACTCAGACATAT 395222 5-10-5 127 10633 10652 0 GGTGGTAATTTGTCACAGCA 395223 5-10-5 84 11308 11327 0 AAGGTCACACAGTTAAGAGT 399850 5-10-5 79 11472 11491 0 AAATGCAGGGCTAAAATCAC 399851 5-10-5 88 12715 12734 0 ACTGGATACATTGGCAGACA 399852 5-10-5 111 12928 12947 0 CTAGAGGAACCACTAGATAT 395201 5-10-5 85 13681 13700 0 ACAAATTCCCAGACTCAGCA 399827 5-10-5 100 13746 13765 0 ATCTCAGGACAGGTGAGCAA 399828 5-10-5 116 13760 13779 0 GAGTAGAGATTCTCATCTCA 395202 5-10-5 129 13816 13835 0 GTGCCATCTGAACAGCACCT 399829 5-10-5 117 13828 13847 0 GAGTCTTCTGAAGTGCCATC 399830 5-10-5 81 13903 13922 0 AAGCAGGGCCTCAGGTGGAA 395203 5-10-5 110 13926 13945 0 CCTGGAACCCCTGCAGCCAG 395204 5-10-5 152 13977 13996 0 TTCAGGCAGGTTGCTGCTAG 399831 5-10-5 83 13986 14005 0 AAGGAAGACTTCAGGCAGGT 395205 5-10-5 140 13998 14017 0 TCAGCCAGGCCAAAGGAAGA 399832 5-10-5 137 14112 14131 0 TAGGGAGAGCTCACAGATGC 395206 5-10-5 136 14122 14141 0 TAGGAGAAAGTAGGGAGAGC 395207 5-10-5 132 14179 14198 0 TAAAAGCTGCAAGAGACTCA 395208 5-10-5 139 14267 14286 0 TCAGAGAAAACAGTCACCGA 399833 5-10-5 92 14397 14416 0 AGAGACAGGAAGCTGCAGCT 395209 5-10-5 142 14404 14423 0 TCATTTTAGAGACAGGAAGC 395210 5-10-5 113 14441 14460 0 GAATAACAGTGATGTCTGGC 395211 5-10-5 138 14494 14513 0 TCACAGCTCACCGAGTCTGC 395212 5-10-5 98 14524 14543 0 AGTGTAAAATAAAGCCCCTA 395213 5-10-5 96 14601 14620 0 AGGACCCAAGTCATCCTGCT 395214 5-10-5 124 14631 14650 0 GGCCATCAGCTGGCAATGCT 399834 5-10-5 82 14670 14689 0 AAGGAAAGGGAGGCCTAGAG 395215 5-10-5 133 14675 14694 0 TAGACAAGGAAAGGGAGGCC 395216 5-10-5 103 14681 14700 0 ATTTCATAGACAAGGAAAGG 395217 5-10-5 155 14801 14820 0 CTTATAGTTAACACACAGAA 399835 5-10-5 156 14809 14828 0 AAGTCAACCTTATAGTTAAC 395160 5-10-5 19 14891 14910 0 CTGGTGTCTAGGAGATACAC 399797 5-10-5 20 14896 14915 0 GTATGCTGGTGTCTAGGAGA 395161 5-10-5 21 14916 14935 0 GATTTCCCGGTGGTCACTCT 399798 5-10-5 22 14922 14941 0 GCCCTCGATTTCCCGGTGGT 399799 5-10-5 23 14936 14955 0 GTGACCATGACCCTGCCCTC 395162 5-10-5 24 14946 14965 0 CTCGAAGTCGGTGACCATGA 395163 5-10-5 25 14979 14998 0 GTGGAAGCGGGTCCCGTCCT 395164 5-10-5 26 15264 15283 0 ACCCCTGCCAGGTGGGTGCC 399800 5-10-5 27 15269 15288 0 TGACCACCCCTGCCAGGTGG 395165 5-10-5 28 15298 15317 0 ACCCTTGGCCACGCCGGCAT 395166 5-10-5 29 15334 15353 0 TTGGCAGTTGAGCACGCGCA 399801 5-10-5 30 15339 15358 0 TTCCCTTGGCAGTTGAGCAC 399853 5-10-5 86 15471 15490 0 ACAGCATTCTTGGTTAGGAG 399854 5-10-5 97 16134 16153 0 AGTCAAGCTGCTGCCCAGAG 399855 5-10-5 120 16668 16687 0 GCTAGTTATTAAGCACCTGC 399856 5-10-5 150 17267 17286 0 TGTGAGCTCTGGCCCAGTGG 399857 5-10-5 115 18377 18396 0 GAGTAAGGCAGGTTACTCTC 399858 5-10-5 134 18408 18427 0 TAGATGTGACTAACATTTAA 395224 5-10-5 157 18561 18580 0 AGGAACAAAGCCAAGGTCAC 395168 5-10-5 32 18593 18612 0 GCTGGCTTTTCCGAATAAAC 395169 5-10-5 33 18705 18724 0 GTGACCAGCACGACCCCAGC 399859 5-10-5 105 19203 19222 0 CACATTAGCCTTGCTCAAGT 399860 5-10-5 151 19913 19932 0 TGTGATGACCTGGAAAGGTG 395170 5-10-5 149 19933 19952 0 TGGGCATTGGTGGCCCCAAC 395171 5-10-5 34 19962 19981 0 CCAAAGTCCCCAGGGTCACC 395172 5-10-5 128 19966 19985 0 GTCCCCAAAGTCCCCAGGGT 399802 5-10-5 35 19971 19990 0 AGTTGGTCCCCAAAGTCCCC 395173 5-10-5 36 19976 19995 0 GCCAAAGTTGGTCCCCAAAG 399803 5-10-5 37 19988 20007 0 GTCCACACAGCGGCCAAAGT 395174 5-10-5 38 19997 20016 0 GGCAAAGAGGTCCACACAGC 395175 5-10-5 39 20025 20044 0 TGGAGGCACCAATGATGTCC 399804 5-10-5 40 20036 20055 0 GCTGCAGTCGCTGGAGGCAC 399805 5-10-5 41 20047 20066 0 ACAAAGCAGGTGCTGCAGTC 399861 5-10-5 158 20100 20119 0 GTGGTGACTTACCAGCCACG 399862 5-10-5 109 20188 20207 0 CCCCTGCACAGAGCCTGGCA 399863 5-10-5 141 20624 20643 0 TCATGGCTGCAATGCCTGGT 399807 5-10-5 43 20635 20654 0 GGCAGACAGCATCATGGCTG 399808 5-10-5 44 20683 20702 0 GAAGTGGATCAGTCTCTGCC 395177 5-10-5 45 20691 20710 0 TTGGCAGAGAAGTGGATCAG 399809 5-10-5 46 20696 20715 0 CATCTTTGGCAGAGAAGTGG 399810 5-10-5 47 20702 20721 0 TGATGACATCTTTGGCAGAG 399811 5-10-5 48 20709 20728 0 GCCTCATTGATGACATCTTT 399812 5-10-5 49 20721 20740 0 TCAGGGAACCAGGCCTCATT 395178 5-10-5 50 20726 20745 0 GGTCCTCAGGGAACCAGGCC 395179 5-10-5 87 20733 20752 0 ACCCGCTGGTCCTCAGGGAA 399813 5-10-5 51 20740 20759 0 GGTCAGTACCCGCTGGTCCT 395180 5-10-5 119 20762 20781 0 GCAGGGCGGCCACCAGGTTG 399864 5-10-5 93 20995 21014 0 AGAGAGGAGGGCTTAAAGAA 399865 5-10-5 95 21082 21101 0 AGCTGCCAACCTGCAAAAAG 399814 5-10-5 53 21091 21110 0 CTGCAAAACAGCTGCCAACC 395182 5-10-5 54 21121 21140 0 GTAGGCCCCGAGTGTGCTGA 399815 5-10-5 55 21186 21205 0 AGAAACTGGAGCAGCTCAGC 399816 5-10-5 56 21192 21211 0 TCCTGGAGAAACTGGAGCAG 399866 5-10-5 143 21481 21500 0 TGAAAATCCATCCAGCACTG 399867 5-10-5 89 21589 21608 0 AGAACCATGGAGCACCTGAG 399868 5-10-5 123 21696 21715 0 GGCACTGCCCTTCCACCAAA 395183 5-10-5 57 22096 22115 0 CGTTGTGGGCCCGGCAGACC 395184 5-10-5 58 22142 22161 0 AGCAGGCAGCACCTGGCAAT 395185 5-10-5 59 22204 22223 0 CACGGGTCCCCATGCTGGCC 395225 5-10-5 126 22292 22311 0 GGTGCATAAGGAGAAAGAGA 395186 5-10-5 60 24095 24114 0 CTTTGCATTCCAGACCTGGG 399817 5-10-5 61 24100 24119 0 CTTGACTTTGCATTCCAGAC 395226 5-10-5 147 24858 24877 0 TGAGTTCATTTAAGAGTGGA 399869 5-10-5 118 24907 24926 0 GCACCATCCAGACCAGAATC 399870 5-10-5 114 25413 25432 0 GAGAGGTTCAGATCCAGGCC 395187 5-10-5 62 26117 26136 0 GGCAGCAGATGGCAACGGCT 395188 5-10-5 154 26217 26236 0 TTTTAAAGCTCAGCCCCAGC 399818 5-10-5 63 26222 26241 0 AACCATTTTAAAGCTCAGCC 395189 5-10-5 64 26311 26330 0 TCAAGGGCCAGGCCAGCAGC 399819 5-10-5 65 26316 26335 0 CCCACTCAAGGGCCAGGCCA 399820 5-10-5 122 26389 26408 0 GGAGGGAGCTTCCTGGCACC 395190 5-10-5 66 26404 26423 0 ATGCCCCACAGTGAGGGAGG 395191 5-10-5 67 26413 26432 0 AATGGTGAAATGCCCCACAG 395192 5-10-5 153 26456 26475 0 TTGGGAGCAGCTGGCAGCAC 395193 5-10-5 68 26557 26576 0 CATGGGAAGAATCCTGCCTC 395194 5-10-5 69 26639 26658 0 GGAGATGAGGGCCATCAGCA 395195 5-10-5 70 26707 26726 0 TAGATGCCATCCAGAAAGCT 399821 5-10-5 71 26713 26732 0 TCTGGCTAGATGCCATCCAG 395196 5-10-5 72 26790 26809 0 GGCATAGAGCAGAGTAAAGG 399822 5-10-5 73 26795 26814 0 AGCCTGGCATAGAGCAGAGT 399823 5-10-5 135 26801 26820 0 TAGCACAGCCTGGCATAGAG 395197 5-10-5 112 27034 27053 0 GAAGAGGCTTGGCTTCAGAG 399824 5-10-5 74 27040 27059 0 AAGTAAGAAGAGGCTTGGCT 395198 5-10-5 75 27244 27263 0 GCTCAAGGAGGGACAGTTGT 395199 5-10-5 76 27279 27298 0 AAAGATAAATGTCTGCTTGC 399825 5-10-5 77 27284 27303 0 ACCCAAAAGATAAATGTCTG 395200 5-10-5 78 27350 27369 0 TCTTCAAGTTACAAAAGCAA 399826 5-10-5 99 27357 27376 0 ATAAATATCTTCAAGTTACA 405861 5-10-5 162 2545 2564 0 AAGGCTAGCACCAGCTCCTC 405862 5-10-5 163 2546 2565 0 CAAGGCTAGCACCAGCTCCT 405863 5-10-5 164 2547 2566 0 GCAAGGCTAGCACCAGCTCC 405864 5-10-5 165 2548 2567 0 CGCAAGGCTAGCACCAGCTC 405865 5-10-5 166 2550 2569 0 AACGCAAGGCTAGCACCAGC 405866 5-10-5 167 2551 2570 0 GAACGCAAGGCTAGCACCAG 405867 5-10-5 168 2552 2571 0 GGAACGCAAGGCTAGCACCA 405868 5-10-5 169 2553 2572 0 CGGAACGCAAGGCTAGCACC 405869 5-10-5 218 14918 14937 0 TCGATTTCCCGGTGGTCACT 405870 5-10-5 219 14919 14938 0 CTCGATTTCCCGGTGGTCAC 405871 5-10-5 220 14920 14939 0 CCTCGATTTCCCGGTGGTCA 405872 5-10-5 221 14921 14940 0 CCCTCGATTTCCCGGTGGTC 405873 5-10-5 222 14923 14942 0 TGCCCTCGATTTCCCGGTGG 405874 5-10-5 223 14924 14943 0 CTGCCCTCGATTTCCCGGTG 405875 5-10-5 224 14925 14944 0 CCTGCCCTCGATTTCCCGGT 405876 5-10-5 225 14926 14945 0 CCCTGCCCTCGATTTCCCGG 405877 5-10-5 246 15294 15313 0 TTGGCCACGCCGGCATCCCG 405878 5-10-5 247 15295 15314 0 CTTGGCCACGCCGGCATCCC 405879 5-10-5 248 15296 15315 0 CCTTGGCCACGCCGGCATCC 405880 5-10-5 249 15297 15316 0 CCCTTGGCCACGCCGGCATC 405881 5-10-5 250 15299 15318 0 CACCCTTGGCCACGCCGGCA 405882 5-10-5 251 15300 15319 0 GCACCCTTGGCCACGCCGGC 405883 5-10-5 252 15301 15320 0 GGCACCCTTGGCCACGCCGG 405884 5-10-5 253 15302 15321 0 TGGCACCCTTGGCCACGCCG 405885 5-10-5 346 20717 20736 0 GGAACCAGGCCTCATTGATG 405886 5-10-5 347 20718 20737 0 GGGAACCAGGCCTCATTGAT 405887 5-10-5 348 20719 20738 0 AGGGAACCAGGCCTCATTGA 405888 5-10-5 349 20720 20739 0 CAGGGAACCAGGCCTCATTG 405889 5-10-5 350 20722 20741 0 CTCAGGGAACCAGGCCTCAT 405890 5-10-5 351 20723 20742 0 CCTCAGGGAACCAGGCCTCA 405891 5-10-5 352 20724 20743 0 TCCTCAGGGAACCAGGCCTC 405892 5-10-5 353 20725 20744 0 GTCCTCAGGGAACCAGGCCT 405893 5-10-5 431 26635 26654 0 ATGAGGGCCATCAGCACCTT 405894 5-10-5 432 26636 26655 0 GATGAGGGCCATCAGCACCT 405895 5-10-5 433 26637 26656 0 AGATGAGGGCCATCAGCACC 405896 5-10-5 434 26638 26657 0 GAGATGAGGGCCATCAGCAC 405897 5-10-5 435 26640 26659 0 TGGAGATGAGGGCCATCAGC 405898 5-10-5 436 26641 26660 0 CTGGAGATGAGGGCCATCAG 405899 5-10-5 437 26642 26661 0 GCTGGAGATGAGGGCCATCA 405900 5-10-5 438 26643 26662 0 AGCTGGAGATGAGGGCCATC 405901 5-10-5 439 26709 26728 0 GCTAGATGCCATCCAGAAAG 405902 5-10-5 440 26710 26729 0 GGCTAGATGCCATCCAGAAA 405903 5-10-5 441 26711 26730 0 TGGCTAGATGCCATCCAGAA 405904 5-10-5 442 26712 26731 0 CTGGCTAGATGCCATCCAGA 405905 5-10-5 443 26714 26733 0 CTCTGGCTAGATGCCATCCA 405906 5-10-5 444 26715 26734 0 CCTCTGGCTAGATGCCATCC 405907 5-10-5 445 26716 26735 0 GCCTCTGGCTAGATGCCATC 405908 5-10-5 446 26717 26736 0 AGCCTCTGGCTAGATGGCAT 405909 5-10-5 267 18595 18614 0 CAGCTGGCTTTTCCGAATAA 405910 5-10-5 268 18597 18616 0 ACCAGCTGGCTTTTCCGAAT 405911 5-10-5 269 18599 18618 0 GGACCAGCTGGCTTTTCCGA 405912 5-10-5 270 18603 18622 0 GGCTGGACCAGCTGGCTTTT 405913 5-10-5 275 18707 18726 0 CGGTGACCAGCACGACCCCA 405914 5-10-5 276 18709 18728 0 AGCGGTGACCAGCACGACCC 405915 5-10-5 277 18711 18730 0 GCAGCGGTGACCAGCACGAC 405916 5-10-5 278 18713 18732 0 CGGCAGCGGTGACCAGCACG 405917 5-10-5 280 18717 18736 0 TTGCCGGCAGCGGTGACCAG 405918 5-10-5 281 18719 18738 0 AGTTGCCGGCAGCGGTGACC 405919 5-10-5 282 18721 18740 0 GAAGTTGCCGGCAGCGGTGA 405920 5-10-5 283 18723 18742 0 CGGAAGTTGCCGGCAGCGGT 405921 5-10-5 284 18725 18744 0 CCCGGAAGTTGCCGGCAGCG 405922 5-10-5 285 18727 18746 0 GTCCCGGAAGTTGCCGGCAG 405923 5-10-5 288 19931 19950 0 GGCATTGGTGGCCCCAACTG 405924 5-10-5 290 19954 19973 0 CCCAGGGTCACCGGCTGGTC 405925 5-10-5 292 19958 19977 0 AGTCCCCAGGGTCACCGGCT 405926 5-10-5 293 19960 19979 0 AAAGTCCCCAGGGTCACCGG 405927 5-10-5 294 19964 19983 0 CCCCAAAGTCCCCAGGGTCA 405928 5-10-5 295 19969 19988 0 TTGGTCCCCAAAGTCCCCAG 405929 5-10-5 296 19973 19992 0 AAAGTTGGTCCCCAAAGTCC 405930 5-10-5 297 19978 19997 0 CGGCCAAAGTTGGTCCCCAA 405931 5-10-5 298 19980 19999 0 AGCGGCCAAAGTTGGTCCCC 405932 5-10-5 299 19982 20001 0 ACAGCGGCCAAAGTTGGTCC 405933 5-10-5 300 19984 20003 0 ACACAGCGGCCAAAGTTGGT 405934 5-10-5 301 19986 20005 0 CCACACAGCGGCCAAAGTTG 405935 5-10-5 302 19990 20009 0 AGGTCCACACAGCGGCCAAA 405936 5-10-5 304 19994 20013 0 AAAGAGGTCCACACAGCGGC 405937 5-10-5 306 20023 20042 0 GAGGCACCAATGATGTCCTC 405938 5-10-5 307 20027 20046 0 GCTGGAGGCACCAATGATGT 405939 5-10-5 308 20029 20048 0 TCGCTGGAGGCACCAATGAT 405940 5-10-5 309 20031 20050 0 AGTCGCTGGAGGCACCAATG 405941 5-10-5 310 20033 20052 0 GCAGTCGCTGGAGGCACCAA 405942 5-10-5 311 20038 20057 0 GTGCTGCAGTCGCTGGAGGC 405943 5-10-5 312 20040 20059 0 AGGTGCTGCAGTCGCTGGAG 405944 5-10-5 314 20043 20062 0 AGCAGGTGCTGCAGTCGCTG 405945 5-10-5 315 20045 20064 0 AAAGCAGGTGCTGCAGTCGC 405946 5-10-5 316 20049 20068 0 ACACAAAGCAGGTGCTGCAG 405947 5-10-5 317 20051 20070 0 TGACACAAAGCAGGTGCTGC 405949 5-10-5 320 20629 20648 0 CAGCATCATGGCTGCAATGC 405950 5-10-5 321 20631 20650 0 GACAGCATCATGGCTGCAAT 405951 5-10-5 322 20633 20652 0 CAGACAGCATCATGGCTGCA 405952 5-10-5 323 20637 20656 0 TCGGCAGACAGCATCATGGC 405953 5-10-5 325 20641 20660 0 CGGCTCGGCAGACAGCATCA 405954 5-10-5 326 20643 20662 0 TCCGGCTCGGCAGACAGCAT 405955 5-10-5 328 20670 20689 0 CTCTGCCTCAACTCGGCCAG 405956 5-10-5 329 20672 20691 0 GTCTCTGCCTCAACTCGGCC 405957 5-10-5 330 20674 20693 0 CAGTCTCTGCCTCAACTCGG 405958 5-10-5 331 20676 20695 0 ATCAGTCTCTGCCTCAACTC 405959 5-10-5 332 20678 20697 0 GGATCAGTCTCTGCCTCAAC 405960 5-10-5 333 20680 20699 0 GTGGATCAGTCTCTGCCTCA 405961 5-10-5 334 20682 20701 0 AAGTGGATCAGTCTCTGCCT 405962 5-10-5 335 20685 20704 0 GAGAAGTGGATCAGTCTCTG 405963 5-10-5 337 20689 20708 0 GGCAGAGAAGTGGATCAGTC 405964 5-10-5 338 20693 20712 0 CTTTGGCAGAGAAGTGGATC 405965 5-10-5 339 20698 20717 0 GACATCTTTGGCAGAGAAGT 405966 5-10-5 340 20700 20719 0 ATGACATCTTTGGCAGAGAA 405967 5-10-5 341 20704 20723 0 ATTGATGACATCTTTGGCAG 405968 5-10-5 342 20706 20725 0 TCATTGATGACATCTTTGGC 405969 5-10-5 343 20711 20730 0 AGGCCTCATTGATGACATCT 405970 5-10-5 344 20713 20732 0 CCAGGCCTCATTGATGACAT 405971 5-10-5 355 20728 20747 0 CTGGTCCTCAGGGAACCAGG 405972 5-10-5 357 20730 20749 0 CGCTGGTCCTCAGGGAACCA 405973 5-10-5 358 20735 20754 0 GTACCCGCTGGTCCTCAGGG 405974 5-10-5 359 20737 20756 0 CAGTACCCGCTGGTCCTCAG 405975 5-10-5 361 21088 21107 0 CAAAACAGCTGCCAACCTGC 405976 5-10-5 362 21093 21112 0 TCCTGCAAAACAGCTGCCAA 405977 5-10-5 363 21095 21114 0 AGTCCTGCAAAACAGCTGCC 405978 5-10-5 365 21118 21137 0 GGCCCCGAGTGTGCTGACCA 405979 5-10-5 366 21123 21142 0 GTGTAGGCCCCGAGTGTGCT 405980 5-10-5 367 21125 21144 0 CCGTGTAGGCCCCGAGTGTG 405981 5-10-5 368 21127 21146 0 ATCCGTGTAGGCCCCGAGTG 405982 5-10-5 370 21131 21150 0 GGCCATCCGTGTAGGCCCCG 405983 5-10-5 371 21133 21152 0 GTGGCCATCCGTGTAGGCCC 405984 5-10-5 372 21181 21200 0 CTGGAGCAGCTCAGCAGCTC 405985 5-10-5 373 21183 21202 0 AACTGGAGCAGCTCAGCAGC 405986 5-10-5 374 21188 21207 0 GGAGAAACTGGAGCAGCTCA 405987 5-10-5 375 21190 21209 0 CTGGAGAAACTGGAGCAGCT 405988 5-10-5 381 22138 22157 0 GGCAGCACCTGGCAATGGCG 405989 5-10-5 383 22140 22159 0 CAGGCAGCACCTGGCAATGG 405990 5-10-5 386 22144 22163 0 GTAGCAGGCAGCACCTGGCA 405991 5-10-5 394 22206 22225 0 GACACGGGTCCCCATGCTGG 405992 5-10-5 396 22208 22227 0 TGGACACGGGTCCCCATGCT 405993 5-10-5 398 22210 22229 0 AGTGGACACGGGTCCCCATG 405994 5-10-5 400 22212 22231 0 GCAGTGGACACGGGTCCCCA 405995 5-10-5 402 22214 22233 0 TGGCAGTGGACACGGGTCCC 405996 5-10-5 412 24097 24116 0 GACTTTGCATTCCAGACCTG 405997 5-10-5 415 24102 24121 0 TCCTTGACTTTGCATTCCAG 405998 5-10-5 426 26120 26139 0 TCCGGCAGCAGATGGCAACG 405999 5-10-5 160 2437 2456 0 GACCGCCTGGAGCTGACGGT 406003 5-10-5 178 6492 6511 0 CAGTGCGCTCTGACTGCGAG 406004 5-10-5 179 6494 6513 0 GGCAGTGCGCTCTGACTGCG 406005 5-10-5 180 6496 6515 0 CGGGCAGTGCGCTCTGACTG 406006 5-10-5 181 6499 6518 0 CGGGGGGCAGTGCGCTCTGA 406007 5-10-5 183 6532 6551 0 AGGTATCCCCGGCGGGCAGC 406008 5-10-5 188 6539 6558 0 CTTGGTGAGGTATCCCCGGC 406009 5-10-5 190 6541 6560 0 ATCTTGGTGAGGTATCCCCG 406010 5-10-5 193 6546 6565 0 GCAGGATCTTGGTGAGGTAT 406011 5-10-5 194 6548 6567 0 ATGCAGGATCTTGGTGAGGT 406012 5-10-5 195 6550 6569 0 ACATGCAGGATCTTGGTGAG 406013 5-10-5 196 6554 6573 0 GAAGACATGCAGGATCTTGG 406014 5-10-5 199 6585 6604 0 TCTTCACCAGGAAGCCAGGA 406015 5-10-5 200 6590 6609 0 ACTCATCTTCACCAGGAAGC 406016 5-10-5 201 6592 6611 0 CCACTCATCTTCACCAGGAA 406017 5-10-5 203 6596 6615 0 GTCGCCACTCATCTTCACCA 406018 5-10-5 204 6598 6617 0 AGGTCGCCACTCATCTTCAC 406019 5-10-5 205 6600 6619 0 GCAGGTCGCCACTCATCTTC 406020 5-10-5 206 6602 6621 0 CAGCAGGTCGCCACTCATCT 406021 5-10-5 210 9207 9226 0 TTCATCCGCCCGGTACCGTG 406022 5-10-5 211 9209 9228 0 TATTCATCCGCCCGGTACCG 406023 5-10-5 212 9212 9231 0 TGGTATTCATCCGCCCGGTA 406024 5-10-5 213 9214 9233 0 GCTGGTATTCATCCGCCCGG 406025 5-10-5 216 14888 14907 0 GTGTCTAGGAGATACACCTC 406026 5-10-5 217 14893 14912 0 TGCTGGTGTCTAGGAGATAC 406027 5-10-5 226 14930 14949 0 ATGACCCTGCCCTCGATTTC 406028 5-10-5 227 14932 14951 0 CCATGACCCTGCCCTCGATT 406029 5-10-5 228 14934 14953 0 GACCATGACCCTGCCCTCGA 406030 5-10-5 229 14938 14957 0 CGGTGACCATGACCCTGCCC 406031 5-10-5 230 14940 14959 0 GTCGGTGACCATGACCCTGC 406032 5-10-5 232 14944 14963 0 CGAAGTCGGTGACCATGACC 406033 5-10-5 237 15261 15280 0 CCTGCCAGGTGGGTGCCATG 406034 5-10-5 238 15266 15285 0 CCACCCCTGCCAGGTGGGTG 406035 5-10-5 239 15271 15290 0 GCTGACCACCCCTGCCAGGT 406036 5-10-5 241 15283 15302 0 GGCATCCCGGCCGCTGACCA 406037 5-10-5 242 15286 15305 0 GCCGGCATCCCGGCCGCTGA 406038 5-10-5 245 15293 15312 0 TGGCCACGCCGGCATCCCGG 406039 5-10-5 257 15330 15349 0 CAGTTGAGCACGCGCAGGCT 406040 5-10-5 258 15332 15351 0 GGCAGTTGAGCACGCGCAGG 406041 5-10-5 259 15336 15355 0 CCTTGGCAGTTGAGCACGCG 406042 5-10-5 260 15341 15360 0 CCTTCCCTTGGCAGTTGAGC 406043 5-10-5 261 15345 15364 0 GTGCCCTTCCCTTGGCAGTT 406044 5-10-5 262 15347 15366 0 CCGTGCCCTTCCCTTGGCAG 406045 5-10-5 266 18591 18610 0 TGGCTTTTCCGAATAAACTC 406478 5-10-5 448 21692 21711 0 CTGCCCTTCCACCAAAATGC 406479 5-10-5 449 21693 21712 0 ACTGCCCTTCCACCAAAATG 406480 5-10-5 450 21694 21713 0 CACTGCCCTTCCACCAAAAT 406481 5-10-5 451 21695 21714 0 GCACTGCCCTTCCACCAAAA 406482 5-10-5 452 21697 21716 0 GGGCACTGCCCTTCCACCAA 406483 5-10-5 453 21698 21717 0 TGGGCACTGCCCTTCCACCA 406484 5-10-5 454 21699 21718 0 CTGGGCACTGCCCTTCCACC 406485 5-10-5 455 21700 21719 0 GCTGGGCACTGCCCTTCCAC 408653 5-10-5 354 20727 20746 0 TGGTCCTCAGGGAACCAGGC 409126 5-10-5 447 15298 15317 1 ACCCTTGGTCACGCCGGCAT 410529 5-10-5 184 6534 6553 0 TGAGGTATCCCCGGCGGGCA 410530 5-10-5 185 6535 6554 0 GTGAGGTATCCCCGGCGGGC 410531 5-10-5 186 6536 6555 0 GGTGAGGTATCCCCGGCGGG 410532 5-10-5 187 6538 6557 0 TTGGTGAGGTATCCCCGGCG 410533 5-10-5 189 6540 6559 0 TCTTGGTGAGGTATCCCCGG 410534 5-10-5 191 6542 6561 0 GATCTTGGTGAGGTATCCCC 410535 5-10-5 192 6544 6563 0 AGGATCTTGGTGAGGTATCC 410536 5-10-5 243 15291 15310 0 GCCACGCCGGCATCCCGGCC 410537 5-10-5 244 15292 15311 0 GGCCACGCCGGCATCCCGGC 410538 5-10-5 254 15303 15322 0 CTGGCACCCTTGGCCACGCC 410539 5-10-5 255 15304 15323 0 GCTGGCACCCTTGGCCACGC 410540 5-10-5 356 20729 20748 0 GCTGGTCCTCAGGGAACCAG 410541 5-10-5 376 22133 22152 0 CACCTGGCAATGGCGTAGAC 410542 5-10-5 377 22134 22153 0 GCACCTGGCAATGGCGTAGA 410543 5-10-5 378 22135 22154 0 AGCACCTGGCAATGGCGTAG 410544 5-10-5 379 22136 22155 0 CAGCACCTGGCAATGGCGTA 410545 5-10-5 380 22137 22156 0 GCAGCACCTGGCAATGGCGT 410546 5-10-5 382 22139 22158 0 AGGCAGCACCTGGCAATGGC 410547 5-10-5 384 22141 22160 0 GCAGGCAGCACCTGGCAATG 410548 5-10-5 385 22143 22162 0 TAGCAGGCAGCACCTGGCAA 410549 5-10-5 388 22199 22218 0 GTCCCCATGCTGGCCTCAGC 410550 5-10-5 389 22200 22219 0 GGTCCCCATGCTGGCCTCAG 410551 5-10-5 390 22201 22220 0 GGGTCCCCATGCTGGCCTCA 410552 5-10-5 391 22202 22221 0 CGGGTCCCCATGCTGGCCTC 410553 5-10-5 392 22203 22222 0 ACGGGTCCCCATGCTGGCCT 410554 5-10-5 393 22205 22224 0 ACACGGGTCCCCATGCTGGC 410555 5-10-5 395 22207 22226 0 GGACACGGGTCCCCATGCTG 410556 5-10-5 397 22209 22228 0 GTGGACACGGGTCCCCATGC 410557 5-10-5 399 22211 22230 0 CAGTGGACACGGGTCCCCAT 410558 5-10-5 401 22213 22232 0 GGCAGTGGACACGGGTCCCC 410559 5-10-5 403 22215 22234 0 GTGGCAGTGGACACGGGTCC 410560 5-10-5 404 22216 22235 0 GGTGGCAGTGGACACGGGTC 410561 5-10-5 405 22217 22236 0 TGGTGGCAGTGGACACGGGT 410562 5-10-5 411 24096 24115 0 ACTTTGCATTCCAGACCTGG 410563 5-10-5 413 24098 24117 0 TGACTTTGCATTCCAGACCT 410564 5-10-5 414 24099 24118 0 TTGACTTTGCATTCCAGACC 410565 5-10-5 419 26112 26131 0 CAGATGGCAACGGCTGTCAC 410566 5-10-5 420 26113 26132 0 GCAGATGGCAACGGCTGTCA 410567 5-10-5 421 26114 26133 0 AGCAGATGGCAAGGGCTGTC 410568 5-10-5 422 26115 26134 0 CAGCAGATGGCAACGGCTGT 410569 5-10-5 423 26116 26135 0 GCAGCAGATGGCAACGGCTG 410570 5-10-5 424 26118 26137 0 CGGCAGCAGATGGCAACGGC 410571 5-10-5 425 26119 26138 0 CCGGCAGCAGATGGCAACGG 410572 5-10-5 427 26121 26140 0 CTCCGGCAGCAGATGGCAAC 410573 5-10-5 428 26122 26141 0 GCTCCGGCAGCAGATGGCAA 410730 5-10-5 202 6594 6613 0 CGCCACTCATCTTCACCAGG 410731 5-10-5 207 6604 6623 0 TCCAGCAGGTCGCCACTCAT 410732 5-10-5 214 9216 9235 0 GGGCTGGTATTCATCCGCCC 410733 5-10-5 231 14942 14961 0 AAGTCGGTGACCATGACCCT 410734 5-10-5 279 18714 18733 0 CCGGCAGCGGTGACCAGCAC 410735 5-10-5 291 19956 19975 0 TCCCCAGGGTCACCGGCTGG 410736 5-10-5 303 19992 20011 0 AGAGGTCCACACAGCGGCCA 410737 5-10-5 313 20042 20061 0 GCAGGTGCTGCAGTCGCTGG 410738 5-10-5 324 20639 20658 0 GCTCGGCAGACAGCATCATG 410739 5-10-5 336 20687 20706 0 CAGAGAAGTGGATCAGTCTC 410740 5-10-5 345 20715 20734 0 AACCAGGCCTCATTGATGAC 410741 5-10-5 369 21129 21148 0 CCATCCGTGTAGGCCCCGAG 410742 5-10-5 159 2433 2452 0 GCCTGGAGCTGACGGTGCCC 410743 5-10-5 170 2560 2579 0 TCCTCCTCGGAACGCAAGGC 410744 5-10-5 171 2585 2604 0 GTGCTCGGGTGCTTCGGCCA 410745 5-10-5 172 2605 2624 0 TGGAAGGTGGCTGTGGTTCC 410746 5-10-5 176 6444 6463 0 CGTAGGTGCCAGGCAACCTC 410747 5-10-5 177 6482 6501 0 TGACTGCGAGAGGTGGGTCT 410748 5-10-5 182 6528 6547 0 ATCCCCGGCGGGCAGCCTGG 410749 5-10-5 197 6565 6584 0 AGAAGGCCATGGAAGACATG 410750 5-10-5 198 6575 6594 0 GAAGCCAGGAAGAAGGCCAT 410752 5-10-5 209 9149 9168 0 GCAAAGACAGAGGAGTCCTC 410753 5-10-5 215 14877 14896 0 ATACACCTCCACCAGGCTGC 410754 5-10-5 233 14954 14973 0 GGCACATTCTCGAAGTCGGT 410756 5-10-5 235 15254 15273 0 GGTGGGTGCCATGACTGTCA 410757 5-10-5 240 15279 15298 0 TCCCGGCCGCTGACCACCCC 410758 5-10-5 256 15309 15328 0 CGCATGCTGGCACCCTTGGC 410759 5-10-5 263 15358 15377 0 GGTGCCGCTAACCGTGCCCT 410761 5-10-5 271 18614 18633 0 GTGGCCCCACAGGCTGGACC 410762 5-10-5 272 18627 18646 0 AGCAGCACCACCAGTGGCCC 410763 5-10-5 273 18649 18668 0 GCTGTACCCACCCGCCAGGG 410764 5-10-5 274 18695 18714 0 CGACCCCAGCCCTCGCCAGG 410767 5-10-5 289 19941 19960 0 GCTGGTCTTGGGCATTGGTG 410768 5-10-5 305 20016 20035 0 CAATGATGTCCTCCCCTGGG 410769 5-10-5 318 20061 20080 0 TCCCACTCTGTGACACAAAG 410770 5-10-5 327 20657 20676 0 CGGCCAGGGTGAGCTCCGGC 410771 5-10-5 360 20785 20804 0 ACCTGCCCCATGGGTGCTGG 410772 5-10-5 364 21106 21125 0 GCTGACCATACAGTCCTGCA 410773 5-10-5 387 22189 22208 0 TGGCCTCAGCTGGTGGAGCT 410774 5-10-5 406 22220 22239 0 TGTTGGTGGCAGTGGACACG 410776 5-10-5 408 23985 24004 0 GTGCCAAGGTCCTCCACCTC 410777 5-10-5 409 24005 24024 0 TCAGCACAGGCGGCTTGTGG 410778 5-10-5 410 24035 24054 0 CCACGCACTGGTTGGGCTGA 410779 5-10-5 416 24115 24134 0 CGGGATTCCATGCTCCTTGA 410781 5-10-5 418 25994 26013 0 GGAGGGCACTGCAGCCAGTC 410782 5-10-5 429 26132 26151 0 CCAGGTGCCGGCTCCGGCAG 410783 5-10-5 430 26142 26161 0 GAGGCCTGCGCCAGGTGCCG

In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 2. In certain such embodiments, the nucleotide sequences illustrated in Table 7 have a 3-14-3 gap-widened motif. Table 9 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 2, having a 3-14-3 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 9 Gapmer antisense compounds having a 3-14-3 motif targeted to SEQ ID NO: 2 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ ID Isis No Motif NO NO: 2 NO: 2 Mismatches Sequence (5′-3′) 399871 3-14-3 4 2274 2293 0 GCGCGGAATCCTGGCTGGGA 399872 3-14-3 5 2381 2400 0 GAGGAGACCTAGAGGCCGTG 399873 3-14-3 6 2439 2458 0 AGGACCGCCTGGAGCTGACG 399874 3-14-3 7 2549 2568 0 ACGCAAGGCTAGCACCAGCT 399875 3-14-3 9 2619 2638 0 CCTTGGCGCAGCGGTGGAAG 399876 3-14-3 10 6498 6517 0 GGCGGGCAGTGCGCTCTGAC 399877 3-14-3 11 6537 6556 0 TGGTGAGGTATCCCCGGCGG 399878 3-14-3 14 6557 6576 0 ATGGAAGACATGCAGGATCT 399879 3-14-3 15 6583 6602 0 TTCACCAGGAAGCCAGGAAG 399880 3-14-3 17 9130 9149 0 CCTCGATGTAGTCGACATGG 399881 3-14-3 18 9210 9229 0 GTATTCATCCGCCCGGTACC 399882 3-14-3 19 14891 14910 0 CTGGTGTCTAGGAGATACAC 399883 3-14-3 21 14916 14935 0 GATTTCCCGGTGGTCACTCT 399884 3-14-3 24 14946 14965 0 CTCGAAGTCGGTGACCATGA 399885 3-14-3 25 14979 14998 0 GTGGAAGCGGGTCCCGTCCT 399886 3-14-3 26 15264 15283 0 ACCCCTGCCAGGTGGGTGCC 399887 3-14-3 28 15298 15317 0 ACCCTTGGCCACGCCGGCAT 399888 3-14-3 29 15334 15353 0 TTGGCAGTTGAGCACGCGCA 399890 3-14-3 32 18593 18612 0 GCTGGCTTTTCCGAATAAAC 399891 3-14-3 33 18705 18724 0 GTGACCAGCACGACCCCAGC 399892 3-14-3 149 19933 19952 0 TGGGCATTGGTGGCCCCAAC 399893 3-14-3 34 19962 19981 0 CCAAAGTCCCCAGGGTCACC 399894 3-14-3 128 19966 19985 0 GTCCCCAAAGTCCCCAGGGT 399895 3-14-3 36 19976 19995 0 GCCAAAGTTGGTCCCCAAAG 399896 3-14-3 38 19997 20016 0 GGCAAAGAGGTCCACACAGC 399897 3-14-3 39 20025 20044 0 TGGAGGCACCAATGATGTCC 399899 3-14-3 45 20691 20710 0 TTGGCAGAGAAGTGGATCAG 399900 3-14-3 50 20726 20745 0 GGTCCTCAGGGAACCAGGCC 399901 3-14-3 87 20733 20752 0 ACCCGCTGGTCCTCAGGGAA 399902 3-14-3 119 20762 20781 0 GCAGGGCGGCCACCAGGTTG 399904 3-14-3 54 21121 21140 0 GTAGGCCCCGAGTGTGCTGA 399905 3-14-3 57 22096 22115 0 CGTTGTGGGCCCGGCAGACC 399906 3-14-3 58 22142 22161 0 AGCAGGCAGCACCTGGCAAT 399907 3-14-3 59 22204 22223 0 CACGGGTCCCCATGCTGGCC 399908 3-14-3 60 24095 24114 0 CTTTGCATTCCAGACCTGGG 399909 3-14-3 62 26117 26136 0 GGCAGCAGATGGCAACGGCT 399910 3-14-3 154 26217 26236 0 TTTTAAAGCTCAGCCCCAGC 399911 3-14-3 64 26311 26330 0 TCAAGGGCCAGGCCAGCAGC 399912 3-14-3 66 26404 26423 0 ATGCCCCACAGTGAGGGAGG 399913 3-14-3 67 26413 26432 0 AATGGTGAAATGCCCCACAG 399914 3-14-3 153 26456 26475 0 TTGGGAGCAGCTGGCAGCAC 399915 3-14-3 68 26557 26576 0 CATGGGAAGAATCCTGCCTC 399916 3-14-3 69 26639 26658 0 GGAGATGAGGGCCATCAGCA 399917 3-14-3 70 26707 26726 0 TAGATGCCATCCAGAAAGCT 399918 3-14-3 72 26790 26809 0 GGCATAGAGCAGAGTAAAGG 399919 3-14-3 112 27034 27053 0 GAAGAGGCTTGGCTTCAGAG 399920 3-14-3 75 27244 27263 0 GCTCAAGGAGGGACAGTTGT 399921 3-14-3 76 27279 27298 0 AAAGATAAATGTCTGCTTGC 399922 3-14-3 78 27350 27369 0 TCTTCAAGTTACAAAAGCAA 399923 3-14-3 85 13681 13700 0 ACAAATTCCCAGACTCAGCA 399924 3-14-3 129 13816 13835 0 GTGCCATCTGAACAGCACCT 399925 3-14-3 110 13926 13945 0 CCTGGAACCCCTGCAGCCAG 399926 3-14-3 152 13977 13996 0 TTCAGGCAGGTTGCTGCTAG 399927 3-14-3 140 13998 14017 0 TCAGCCAGGCCAAAGGAAGA 399928 3-14-3 136 14122 14141 0 TAGGAGAAAGTAGGGAGAGC 399929 3-14-3 132 14179 14198 0 TAAAAGCTGCAAGAGACTCA 399930 3-14-3 139 14267 14286 0 TCAGAGAAAACAGTCACCGA 399931 3-14-3 142 14404 14423 0 TCATTTTAGAGACAGGAAGC 399932 3-14-3 113 14441 14460 0 GAATAACAGTGATGTCTGGC 399933 3-14-3 138 14494 14513 0 TCACAGCTCACCGAGTCTGC 399934 3-14-3 98 14524 14543 0 AGTGTAAAATAAAGCCCCTA 399935 3-14-3 96 14601 14620 0 AGGACCCAAGTCATCCTGCT 399936 3-14-3 124 14631 14650 0 GGCCATCAGCTGGCAATGCT 399937 3-14-3 133 14675 14694 0 TAGACAAGGAAAGGGAGGCC 399938 3-14-3 103 14681 14700 0 ATTTCATAGACAAGGAAAGG 399939 3-14-3 155 14801 14820 0 CTTATAGTTAACACACAGAA 399943 3-14-3 102 5590 5609 0 ATGTGCAGAGATCAATCACA 399944 3-14-3 127 10633 10652 0 GGTGGTAATTTGTCACAGCA 399945 3-14-3 84 11308 11327 0 AAGGTCACACAGTTAAGAGT 399946 3-14-3 157 18561 18580 0 AGGAACAAAGCCAAGGTCAC 399947 3-14-3 126 22292 22311 0 GGTGCATAAGGAGAAAGAGA 399948 3-14-3 147 24858 24877 0 TGAGTTCATTTAAGAGTGGA 399949 3-14-3 8 2556 2575 0 CCTCGGAACGCAAGGCTAGC 399950 3-14-3 12 6543 6562 0 GGATCTTGGTGAGGTATCCC 399951 3-14-3 13 6552 6571 0 AGACATGCAGGATCTTGGTG 399952 3-14-3 16 6588 6607 0 TCATCTTCACCAGGAAGCCA 399953 3-14-3 20 14896 14915 0 GTATGCTGGTGTCTAGGAGA 399954 3-14-3 22 14922 14941 0 GCCCTCGATTTCCCGGTGGT 399955 3-14-3 23 14936 14955 0 GTGACCATGACCCTGCCCTC 399956 3-14-3 27 15269 15288 0 TGACCACCCCTGCCAGGTGG 399957 3-14-3 30 15339 15358 0 TTCCCTTGGCAGTTGAGCAC 399958 3-14-3 35 19971 19990 0 AGTTGGTCCCCAAAGTCCCC 399959 3-14-3 37 19988 20007 0 GTCCACACAGCGGCCAAAGT 399960 3-14-3 40 20036 20055 0 GCTGCAGTCGCTGGAGGCAC 399961 3-14-3 41 20047 20066 0 ACAAAGCAGGTGCTGCAGTC 399963 3-14-3 43 20635 20654 0 GGCAGACAGCATCATGGCTG 399964 3-14-3 44 20683 20702 0 GAAGTGGATCAGTCTCTGCC 399965 3-14-3 46 20696 20715 0 CATCTTTGGCAGAGAAGTGG 399966 3-14-3 47 20702 20721 0 TGATGACATCTTTGGCAGAG 399967 3-14-3 48 20709 20728 0 GCCTCATTGATGACATCTTT 399968 3-14-3 49 20721 20740 0 TCAGGGAACCAGGCCTCATT 399969 3-14-3 51 20740 20759 0 GGTCAGTACCCGCTGGTCCT 399970 3-14-3 53 21091 21110 0 CTGCAAAACAGCTGCCAACC 399971 3-14-3 55 21186 21205 0 AGAAACTGGAGCAGCTCAGC 399972 3-14-3 56 21192 21211 0 TCCTGGAGAAACTGGAGCAG 399973 3-14-3 61 24100 24119 0 CTTGACTTTGCATTCCAGAC 399974 3-14-3 63 26222 26241 0 AACCATTTTAAAGCTCAGCC 399975 3-14-3 65 26316 26335 0 CCCACTCAAGGGCCAGGCCA 399976 3-14-3 122 26389 26408 0 GGAGGGAGCTTCCTGGCACC 399977 3-14-3 71 26713 26732 0 TCTGGCTAGATGCCATCCAG 399978 3-14-3 73 26795 26814 0 AGCCTGGCATAGAGCAGAGT 399979 3-14-3 135 26801 26820 0 TAGCACAGCCTGGCATAGAG 399980 3-14-3 74 27040 27059 0 AAGTAAGAAGAGGCTTGGCT 399981 3-14-3 77 27284 27303 0 ACCCAAAAGATAAATGTCTG 399982 3-14-3 99 27357 27376 0 ATAAATATCTTCAAGTTACA 399983 3-14-3 100 13746 13765 0 ATCTCAGGACAGGTGAGCAA 399984 3-14-3 116 13760 13779 0 GAGTAGAGATTCTCATCTCA 399985 3-14-3 117 13828 13847 0 GAGTCTTCTGAAGTGCCATC 399986 3-14-3 81 13903 13922 0 AAGCAGGGCCTCAGGTGGAA 399987 3-14-3 83 13986 14005 0 AAGGAAGACTTCAGGCAGGT 399988 3-14-3 137 14112 14131 0 TAGGGAGAGCTCACAGATGC 399989 3-14-3 92 14397 14416 0 AGAGACAGGAAGCTGCAGCT 399990 3-14-3 82 14670 14689 0 AAGGAAAGGGAGGCCTAGAG 399991 3-14-3 156 14809 14828 0 AAGTCAACCTTATAGTTAAC 399993 3-14-3 107 3056 3075 0 CCCACTATAATGGCAAGCCC 399994 3-14-3 80 4306 4325 0 AACCCAGTTCTAATGCACCT 399995 3-14-3 106 5140 5159 0 CCAGTCAGAGTAGAACAGAG 399996 3-14-3 121 5599 5618 0 GGAGCCTACATGTGCAGAGA 399997 3-14-3 94 5667 5686 0 AGCATGGCACCAGCATCTGC 399998 3-14-3 108 6652 6671 0 CCCAGCCCTATCAGGAAGTG 399999 3-14-3 144 7099 7118 0 TGACATCCAGGAGGGAGGAG 400000 3-14-3 91 7556 7575 0 AGACTGATGGAAGGCATTGA 400001 3-14-3 131 7565 7584 0 GTGTTGAGCAGACTGATGGA 400002 3-14-3 145 8836 8855 0 TGACATCTTGTCTGGGAGCC 400003 3-14-3 90 8948 8967 0 AGACTAGGAGCCTGAGTTTT 400004 3-14-3 125 9099 9118 0 GGCCTGCAGAAGCCAGAGAG 400005 3-14-3 148 10252 10271 0 TGGCAGCAACTCAGACATAT 400006 3-14-3 79 11472 11491 0 AAATGCAGGGCTAAAATCAC 400007 3-14-3 88 12715 12734 0 ACTGGATACATTGGCAGACA 400008 3-14-3 111 12928 12947 0 CTAGAGGAACCACTAGATAT 400009 3-14-3 86 15471 15490 0 ACAGCATTCTTGGTTAGGAG 400010 3-14-3 97 16134 16153 0 AGTCAAGCTGCTGCCCAGAG 400011 3-14-3 120 16668 16687 0 GCTAGTTATTAAGCACCTGC 400012 3-14-3 150 17267 17286 0 TGTGAGCTCTGGCCCAGTGG 400013 3-14-3 115 18377 18396 0 GAGTAAGGCAGGTTACTCTC 400014 3-14-3 134 18408 18427 0 TAGATGTGACTAACATTTAA 400015 3-14-3 105 19203 19222 0 CACATTAGCCTTGCTCAAGT 400016 3-14-3 151 19913 19932 0 TGTGATGACCTGGAAAGGTG 400017 3-14-3 158 20100 20119 0 GTGGTGACTTACCAGCCACG 400018 3-14-3 109 20188 20207 0 CCCCTGCACAGAGCCTGGCA 400019 3-14-3 141 20624 20643 0 TCATGGCTGCAATGCCTGGT 400020 3-14-3 93 20995 21014 0 AGAGAGGAGGGCTTAAAGAA 400021 3-14-3 95 21082 21101 0 AGCTGCCAACCTGCAAAAAG 400022 3-14-3 143 21481 21500 0 TGAAAATCCATCCAGCACTG 400023 3-14-3 89 21589 21608 0 AGAACCATGGAGCACCTGAG 400024 3-14-3 123 21696 21715 0 GGCACTGCCCTTCCACCAAA 400025 3-14-3 118 24907 24926 0 GCACCATCCAGACCAGAATC 400026 3-14-3 114 25413 25432 0 GAGAGGTTCAGATCCAGGCC 405604 3-14-3 236 15257 15276 0 CCAGGTGGGTGCCATGACTG 405641 3-14-3 373 21183 21202 0 AACTGGAGCAGCTCAGCAGC 410574 3-14-3 184 6534 6553 0 TGAGGTATCCCCGGCGGGCA 410575 3-14-3 185 6535 6554 0 GTGAGGTATCCCCGGCGGGC 410576 3-14-3 186 6536 6555 0 GGTGAGGTATCCCCGGCGGG 410577 3-14-3 187 6538 6557 0 TTGGTGAGGTATCCCCGGCG 410578 3-14-3 188 6539 6558 0 CTTGGTGAGGTATCCCCGGC 410579 3-14-3 189 6540 6559 0 TCTTGGTGAGGTATCCCCGG 410580 3-14-3 190 6541 6560 0 ATCTTGGTGAGGTATCCCCG 410581 3-14-3 191 6542 6561 0 GATCTTGGTGAGGTATCCCC 410582 3-14-3 192 6544 6563 0 AGGATCTTGGTGAGGTATCC 410583 3-14-3 243 15291 15310 0 GCCACGCCGGCATCCCGGCC 410584 3-14-3 244 15292 15311 0 GGCCACGCCGGCATCCCGGC 410585 3-14-3 245 15293 15312 0 TGGCCACGCCGGCATCCCGG 410586 3-14-3 246 15294 15313 0 TTGGCCACGCCGGCATCCCG 410587 3-14-3 247 15295 15314 0 CTTGGCCACGCCGGCATCCC 410588 3-14-3 248 15296 15315 0 CCTTGGCCACGCCGGCATCC 410589 3-14-3 249 15297 15316 0 CCCTTGGCCACGCCGGCATC 410590 3-14-3 250 15299 15318 0 CACCCTTGGCCACGCCGGCA 410591 3-14-3 251 15300 15319 0 GCACCCTTGGCCACGCCGGC 410592 3-14-3 252 15301 15320 0 GGCACCCTTGGCCACGCCGG 410593 3-14-3 253 15302 15321 0 TGGCACCCTTGGCCACGCCG 410594 3-14-3 254 15303 15322 0 CTGGCACCCTTGGCCACGCC 410595 3-14-3 255 15304 15323 0 GCTGGCACCCTTGGCCACGC 410596 3-14-3 348 20719 20738 0 AGGGAACCAGGCCTCATTGA 410597 3-14-3 349 20720 20739 0 CAGGGAACCAGGCCTCATTG 410598 3-14-3 350 20722 20741 0 CTCAGGGAACCAGGCCTCAT 410599 3-14-3 351 20723 20742 0 CCTCAGGGAACCAGGCCTCA 410600 3-14-3 352 20724 20743 0 TCCTCAGGGAACCAGGCCTC 410601 3-14-3 353 20725 20744 0 GTCCTCAGGGAACCAGGCCT 410602 3-14-3 354 20727 20746 0 TGGTCCTCAGGGAACCAGGC 410603 3-14-3 355 20728 20747 0 CTGGTCCTCAGGGAACCAGG 410604 3-14-3 356 20729 20748 0 GCTGGTCCTCAGGGAACCAG 410605 3-14-3 376 22133 22152 0 CACCTGGCAATGGCGTAGAC 410606 3-14-3 377 22134 22153 0 GCACCTGGCAATGGCGTAGA 410607 3-14-3 378 22135 22154 0 AGCACCTGGCAATGGCGTAG 410608 3-14-3 379 22136 22155 0 CAGCACCTGGCAATGGCGTA 410609 3-14-3 380 22137 22156 0 GCAGCACCTGGCAATGGCGT 410610 3-14-3 381 22138 22157 0 GGCAGCACCTGGCAATGGCG 410611 3-14-3 382 22139 22158 0 AGGCAGCACCTGGCAATGGC 410612 3-14-3 383 22140 22159 0 CAGGCAGCACCTGGCAATGG 410613 3-14-3 384 22141 22160 0 GCAGGCAGCACCTGGCAATG 410614 3-14-3 385 22143 22162 0 TAGCAGGCAGCACCTGGCAA 410615 3-14-3 388 22199 22218 0 GTCCCCATGCTGGCCTCAGC 410616 3-14-3 389 22200 22219 0 GGTCCCCATGCTGGCCTCAG 410617 3-14-3 390 22201 22220 0 GGGTCCCCATGCTGGCCTCA 410618 3-14-3 391 22202 22221 0 CGGGTCCCCATGCTGGCCTC 410619 3-14-3 392 22203 22222 0 ACGGGTCCCCATGCTGGCCT 410620 3-14-3 393 22205 22224 0 ACACGGGTCCCCATGCTGGC 410621 3-14-3 394 22206 22225 0 GACACGGGTCCCCATGCTGG 410622 3-14-3 395 22207 22226 0 GGACACGGGTCCCCATGCTG 410623 3-14-3 396 22208 22227 0 TGGACACGGGTCCCCATGCT 410624 3-14-3 397 22209 22228 0 GTGGACACGGGTCCCCATGC 410625 3-14-3 398 22210 22229 0 AGTGGACACGGGTCCCCATG 410626 3-14-3 399 22211 22230 0 CAGTGGACACGGGTCCCCAT 410627 3-14-3 400 22212 22231 0 GCAGTGGACACGGGTCCCCA 410628 3-14-3 401 22213 22232 0 GGCAGTGGACACGGGTCCCC 410629 3-14-3 402 22214 22233 0 TGGCAGTGGACACGGGTCCC 410630 3-14-3 403 22215 22234 0 GTGGCAGTGGACACGGGTCC 410631 3-14-3 404 22216 22235 0 GGTGGCAGTGGACACGGGTC 410632 3-14-3 405 22217 22236 0 TGGTGGCAGTGGACACGGGT 410633 3-14-3 411 24096 24115 0 ACTTTGCATTCCAGACCTGG 410634 3-14-3 412 24097 24116 0 GACTTTGCATTCCAGACCTG 410635 3-14-3 413 24098 24117 0 TGACTTTGCATTCCAGACCT 410636 3-14-3 414 24099 24118 0 TTGACTTTGCATTCCAGACC 410637 3-14-3 419 26112 26131 0 CAGATGGCAACGGCTGTCAC 410638 3-14-3 420 26113 26132 0 GCAGATGGCAACGGCTGTCA 410639 3-14-3 421 26114 26133 0 AGCAGATGGCAACGGCTGTC 410640 3-14-3 422 26115 26134 0 CAGCAGATGGCAACGGCTGT 410641 3-14-3 423 26116 26135 0 GCAGCAGATGGCAACGGCTG 410642 3-14-3 424 26118 26137 0 CGGCAGCAGATGGCAACGGC 410643 3-14-3 425 26119 26138 0 CCGGCAGCAGATGGCAACGG 410644 3-14-3 426 26120 26139 0 TCCGGCAGCAGATGGCAACG 410645 3-14-3 427 26121 26140 0 CTCCGGCAGCAGATGGCAAC 410646 3-14-3 428 26122 26141 0 GCTCCGGCAGCAGATGGCAA

In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 2. In certain such embodiments, the nucleotide sequences illustrated in Table 7 have a 2-13-5 gap-widened motif. Table 10 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 2, having a 2-13-5 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 10 Gapmer antisense compounds having a 2-13-5 motif targeted to SEQ ID NO: 2 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ ID Isis No Motif NO NO: 2 NO: 2 Mismatches Sequence (5′-3′) 410647 2-13-5 184 6534 6553 0 TGAGGTATCCCCGGCGGGCA 410648 2-13-5 185 6535 6554 0 GTGAGGTATCCCCGGCGGGC 410649 2-13-5 186 6536 6555 0 GGTGAGGTATCCCCGGCGGG 410650 2-13-5 11 6537 6556 0 TGGTGAGGTATCCCCGGCGG 410651 2-13-5 187 6538 6557 0 TTGGTGAGGTATCCCCGGCG 410652 2-13-5 188 6539 6558 0 CTTGGTGAGGTATCCCCGGC 410653 2-13-5 189 6540 6559 0 TCTTGGTGAGGTATCCCCGG 410654 2-13-5 190 6541 6560 0 ATCTTGGTGAGGTATCCCCG 410655 2-13-5 191 6542 6561 0 GATCTTGGTGAGGTATCCCC 410656 2-13-5 12 6543 6562 0 GGATCTTGGTGAGGTATCCC 410657 2-13-5 192 6544 6563 0 AGGATCTTGGTGAGGTATCC 410658 2-13-5 243 15291 15310 0 GCCACGCCGGCATCCCGGCC 410659 2-13-5 244 15292 15311 0 GGCCACGCCGGCATCCCGGC 410660 2-13-5 245 15293 15312 0 TGGCCACGCCGGCATCCCGG 410661 2-13-5 246 15294 15313 0 TTGGCCACGCCGGCATCCCG 410662 2-13-5 247 15295 15314 0 CTTGGCCACGCCGGCATCCC 410663 2-13-5 248 15296 15315 0 CCTTGGCCACGCCGGCATCC 410664 2-13-5 249 15297 15316 0 CCCTTGGCCACGCCGGCATC 410665 2-13-5 28 15298 15317 0 ACCCTTGGCCACGCCGGCAT 410666 2-13-5 250 15299 15318 0 CACCCTTGGCCACGCCGGCA 410667 2-13-5 251 15300 15319 0 GCACCCTTGGCCACGCCGGC 410668 2-13-5 252 15301 15320 0 GGCACCCTTGGCCACGCCGG 410669 2-13-5 253 15302 15321 0 TGGCACCCTTGGCCACGCCG 410670 2-13-5 254 15303 15322 0 CTGGCACCCTTGGCCACGCC 410671 2-13-5 255 15304 15323 0 GCTGGCACCCTTGGCCACGC 410672 2-13-5 348 20719 20738 0 AGGGAACCAGGCCTCATTGA 410673 2-13-5 349 20720 20739 0 CAGGGAACCAGGCCTCATTG 410674 2-13-5 49 20721 20740 0 TCAGGGAACCAGGCCTCATT 410675 2-13-5 350 20722 20741 0 CTCAGGGAACCAGGCCTCAT 410676 2-13-5 351 20723 20742 0 CCTCAGGGAACCAGGCCTCA 410677 2-13-5 352 20724 20743 0 TCCTCAGGGAACCAGGCCTC 410678 2-13-5 353 20725 20744 0 GTCCTCAGGGAACCAGGCCT 410679 2-13-5 50 20726 20745 0 GGTCCTCAGGGAACCAGGCC 410680 2-13-5 354 20727 20746 0 TGGTCCTCAGGGAACCAGGC 410681 2-13-5 355 20728 20747 0 CTGGTCCTCAGGGAACCAGG 410682 2-13-5 356 20729 20748 0 GCTGGTCCTCAGGGAACCAG 410683 2-13-5 376 22133 22152 0 CACCTGGCAATGGCGTAGAC 410684 2-13-5 377 22134 22153 0 GCACCTGGCAATGGCGTAGA 410685 2-13-5 378 22135 22154 0 AGCACCTGGCAATGGCGTAG 410686 2-13-5 379 22136 22155 0 CAGCACCTGGCAATGGCGTA 410687 2-13-5 380 22137 22156 0 GCAGCACCTGGCAATGGCGT 410688 2-13-5 381 22138 22157 0 GGCAGCACCTGGCAATGGCG 410689 2-13-5 382 22139 22158 0 AGGCAGCACCTGGCAATGGC 410690 2-13-5 383 22140 22159 0 CAGGCAGCACCTGGCAATGG 410691 2-13-5 384 22141 22160 0 GCAGGCAGCACCTGGCAATG 410692 2-13-5 58 22142 22161 0 AGCAGGCAGCACCTGGCAAT 410693 2-13-5 385 22143 22162 0 TAGCAGGCAGCACCTGGCAA 410694 2-13-5 388 22199 22218 0 GTCCCCATGCTGGCCTCAGC 410695 2-13-5 389 22200 22219 0 GGTCCCCATGCTGGCCTCAG 410696 2-13-5 390 22201 22220 0 GGGTCCCCATGCTGGCCTCA 410697 2-13-5 391 22202 22221 0 CGGGTCCCCATGCTGGCCTC 410698 2-13-5 392 22203 22222 0 ACGGGTCCCCATGCTGGCCT 410699 2-13-5 59 22204 22223 0 CACGGGTCCCCATGCTGGCC 410700 2-13-5 393 22205 22224 0 ACACGGGTCCCCATGCTGGC 410701 2-13-5 394 22206 22225 0 GACACGGGTCCCCATGCTGG 410702 2-13-5 395 22207 22226 0 GGACACGGGTCCCCATGCTG 410703 2-13-5 396 22208 22227 0 TGGACACGGGTCCCCATGCT 410704 2-13-5 397 22209 22228 0 GTGGACACGGGTCCCCATGC 410705 2-13-5 398 22210 22229 0 AGTGGACACGGGTCCCCATG 410706 2-13-5 399 22211 22230 0 CAGTGGACACGGGTCCCCAT 410707 2-13-5 400 22212 22231 0 GCAGTGGACACGGGTCCCCA 410708 2-13-5 401 22213 22232 0 GGCAGTGGACACGGGTCCCC 410709 2-13-5 402 22214 22233 0 TGGCAGTGGACACGGGTCCC 410710 2-13-5 403 22215 22234 0 GTGGCAGTGGACACGGGTCC 410711 2-13-5 404 22216 22235 0 GGTGGCAGTGGACACGGGTC 410712 2-13-5 405 22217 22236 0 TGGTGGCAGTGGACACGGGT 410713 2-13-5 60 24095 24114 0 CTTTGCATTCCAGACCTGGG 410714 2-13-5 411 24096 24115 0 ACTTTGCATTCCAGACCTGG 410715 2-13-5 412 24097 24116 0 GACTTTGCATTCCAGACCTG 410716 2-13-5 413 24098 24117 0 TGACTTTGCATTCCAGACCT 410717 2-13-5 414 24099 24118 0 TTGACTTTGCATTCCAGACC 410718 2-13-5 61 24100 24119 0 CTTGACTTTGCATTCCAGAC 410719 2-13-5 419 26112 26131 0 CAGATGGCAACGGCTGTCAC 410720 2-13-5 420 26113 26132 0 GCAGATGGCAACGGCTGTCA 410721 2-13-5 421 26114 26133 0 AGCAGATGGCAACGGCTGTC 410722 2-13-5 422 26115 26134 0 CAGCAGATGGCAACGGCTGT 410723 2-13-5 423 26116 26135 0 GCAGCAGATGGCAACGGCTG 410724 2-13-5 62 26117 26136 0 GGCAGCAGATGGCAACGGCT 410725 2-13-5 424 26118 26137 0 CGGCAGCAGATGGCAACGGC 410726 2-13-5 425 26119 26138 0 CCGGCAGCAGATGGCAACGG 410727 2-13-5 426 26120 26139 0 TCCGGCAGCAGATGGCAACG 410728 2-13-5 427 26121 26140 0 CTCCGGCAGCAGATGGCAAC 410729 2-13-5 428 26122 26141 0 GCTCCGGCAGCAGATGGCAA

In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 2. In certain such embodiments, the nucleotide sequences illustrated in Table 7 have a 3-13-4 gap-widened motif. Table 11 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 2, having a 3-13-4 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 11 Gapmer antisense compounds having a 3-13-4 motif targeted to SEQ ID NO: 2 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ ID Isis No Motif NO NO: 2 NO: 2 Mismatches Sequence (5′-3′) 405526 3-13-4 236 15257 15276 0 CCAGGTGGGTGCCATGACTG 405557 3-13-4 50 20726 20745 0 GGTCCTCAGGGAACCAGGCC 405564 3-13-4 373 21183 21202 0 AACTGGAGCAGCTCAGCAGC

The following embodiments set forth target regions of PCSK9 nucleic acids. Also illustrated are examples of antisense compounds targeted to the target regions. It is understood that the sequence set forth in each SEQ ID NO is independent of any modification to a sugar moiety, an internucleoside linkage, or a nucleobase. As such, antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an internucleoside linkage, or a nucleobase. Antisense compounds described by Isis Number (Isis No) indicate a combination of nucleobase sequence and motif.

In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 2: 2274-2400, 2274-2575, 2433-2570, 2433-2579, 2549-2575, 2552-2579, 2585-2638, 2605-2638, 3056-3075, 4150-5159, 4306-4325, 5590-5618, 5667-5686, 6444-6463, 6482-6518, 6492-6518, 6528-6555, 6528-6623, 6534-6561, 6535-6562, 6536-6563, 6537-6563, 6538-6565, 6539-6565, 6540-6567, 6541-6567, 6542-6569, 6546-6573, 6557-6584, 6575-6602, 6585-6611, 6594-6621, 6596-6623, 6652-6671, 7099-7118, 7556-7584, 8836-8855, 8948-8967, 9099-9118, 9099-9168, 9130-9168, 9207-9233, 9207-9235, 9209-9235, 10252-10271, 10633-10652, 11308-11491, 12715-12734, 12928-12947, 13681-13700, 13746-13779, 13816-13847, 13903-13945, 13977-14141, 14179-14198, 14267-14286, 14397-14423, 14441-14460, 14494-14513, 14494-14543, 14524-14543, 14601-14650, 14670-14700, 14675-14700, 14801-14828, 14877-14912, 14877-14915, 14877-14973, 14916-14943, 14916-14973, 14925-14951, 14934-14963, 14946-14973, 14979-14998, 15254-15280, 15254-15328, 15264-15290, 15279-15305, 15291-15318, 15292-15319, 15293-15320, 15294-15321, 15294-15321, 15295-15322, 15296-15315, 15296-15323, 15297-15323, 15298-15323, 15299-15323, 15300-15323, 15301-15328, 15330-15355, 15330-15490, 15339-15366, 15358-15490, 16134-16153, 16668-16687, 17267-17286, 18377-18427, 18561-18580, 18591-18618, 18591-18646, 18591-18668, 18695-18746, 18705-18730, 18709-18736, 18719-18746, 19203-20080, 19931-19952, 19954-19981, 19964-19990, 19973-19999, 19982-20009, 19992-20016, 20016-20042, 20025-20052, 20036-20062, 20045-20070, 20100-20119, 20188-20207, 20624-20650, 20624-20759, 20629-20804, 20633-20660, 20635-20781, 20643-20662, 20657-20676, 20670-20697, 20680-20706, 20683-20781, 20689-20715, 20698-20725, 20709-20736, 20717-20744, 20718-20745, 20719-20746, 20720-20747, 20721-20748, 20722-20749, 20727-20752, 20735-20759, 20762-21014, 20785-21014, 21082-21107, 21082-21152, 21091-21114, 21118-21144, 21127-21152, 21181-21209, 21181-21211, 21183-21211, 21481-21500, 21589-21608, 21692-21719, 22000-22227, 22096-22115, 22096-22223, 22096-22311, 22133-22160, 22133-22163, 22134-22161, 22135-22162, 22136-22163, 22137-22163, 22138-22163, 22189-22239, 22199-22226, 22199-22227, 22200-22227, 22201-22228, 22202-22229, 22203-22230, 22204-22231, 22205-22232, 22206-22233, 22207-22234, 22208-22235, 22209-22236, 22210-22236, 22210-22239, 22211-22236, 22212-22239, 22292-22311, 23985-24054, 24035-24134, 24095-24121, 24858-24877, 24907-24926, 25413-25432, 25994-26013, 26112-26139, 26112-26161, 26112-27303, 26113-26140, 26114-26141, 26115-26141, 26116-26141, 26117-26141, 26117-26475, 26118-26141, 26120-26141, 26132-26151, 26142-26161, 26217-26241, 26311-26335, 26389-26432, 26456-26576, 26635-26662, 26707-26734, 26707-26736, 26790-26820, 27034-27263, 27279-27303, or 27350-27376.

In certain embodiments, a target region is nucleotides 2274-2400 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2274-2400 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NQs: 4 or 5. In certain such embodiments, an antisense compound targeted to nucleotides 2274-2400 of SEQ ID NO: 2 is selected from ISIS NOs: 395149, 399871, 395150 or 399872.

In certain embodiments, a target region is nucleotides 2274-2575 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2274-2575 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 4, 5, 6, 7, 8, 159, 160, 162, 163, 164, 165, 166, 167, 168 or 169. In certain such embodiments, an antisense compound targeted to nucleotides 2274-2575 of SEQ ID NO: 2 is selected from ISIS NOs: 395149, 399871, 395150, 399872, 410742, 405999, 395151, 399873, 405861, 405862, 405863, 405864, 395152, 399874, 405865, 405866, 405867, 405868, 399793, or 399949.

In certain embodiments, a target region is nucleotides 2433-2570 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2433-2570 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 6, 7, 159, 160, 162, 163, 164, 165, 166, or 167. In certain such embodiments, an antisense compound targeted to nucleotides 2433-2570 of SEQ ID NO: 2 is selected from ISIS NOs: 395151, 395152, 399873, 399874, 405861, 405862, 405863, 405864, 405865, 405866, 405999, or 410742.

In certain embodiments, a target region is nucleotides 2433-2579 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2433-2579 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 6, 7, 8, 159, 160, 162, 163, 164, 165, 166, 167, 168, 169, or 170. In certain such embodiments, an antisense compound targeted to nucleotides 2433-2579 of SEQ ID NO: 2 is selected from ISIS NOs: 395151, 395152, 399793, 399873, 399874, 399949, 405861, 405862, 405863, 405864, 405865, 405866, 405867, 405868, 405999, 410742, or 410743.

In certain embodiments, a target region is nucleotides 2549-2575 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2549-2575 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 7, 8, 166, 167, 168 or 169. In certain such embodiments, an antisense compound targeted to nucleotides 2549-2575 of SEQ ID NO: 2 is selected from ISIS NOs: 395152, 399793, 399874, 399949, 405865, 405866, 405867, or 405868.

In certain embodiments, a target region is nucleotides 2552-2579 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2552-2579 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 8, 168, 169 or 170. In certain such embodiments, an antisense compound targeted to nucleotides 2552-2579 of SEQ ID NO: 2 is selected from ISIS NOs: 399793, 399949, 405867, 405868, or 410743.

In certain embodiments, a target region is nucleotides 2585-2638 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2585-2638 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 9, 171 or 172. In certain such embodiments, an antisense compound targeted to nucleotides 2585-2638 of SEQ ID NO: 2 is selected from ISIS NOs: 395153, 399875, 410744, or 410745.

In certain embodiments, a target region is nucleotides 2605-2638 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2605-2638 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 8, 168 or 169. In certain such embodiments, an antisense compound targeted to nucleotides 2605-2638 of SEQ ID NO: 2 is selected from ISIS NOs: 410745, 395153, or 399875.

In certain embodiments, a target region is nucleotides 3056-3075 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 3056-3075 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 107. In certain such embodiments, an antisense compound targeted to nucleotides 3056-3075 of SEQ ID NO: 2 is selected from ISIS NOs: 399837 or 399993.

In certain embodiments, a target region is nucleotides 4150-5159 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 4150-5159 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 106. In certain such embodiments, an antisense compound targeted to nucleotides 4150-5159 of SEQ ID NO: 2 is selected from ISIS NOs: 399839 or 399995.

In certain embodiments, a target region is nucleotides 4306-4325 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 4306-4325 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 80. In certain such embodiments, an antisense compound targeted to nucleotides 4306-4325 of SEQ ID NO: 2 is selected from ISIS NOs: 399838 or 399994.

In certain embodiments, a target region is nucleotides 5590-5618 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 5590-5618 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 102 or 121. In certain such embodiments, an antisense compound targeted to nucleotides 5590-5618 of SEQ ID NO: 2 is selected from ISIS NOs: 395221, 399840, 399943, or 399996.

In certain embodiments, a target region is nucleotides 5667-5686 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 5667-5686 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 94. In certain such embodiments, an antisense compound targeted to nucleotides 5667-5686 of SEQ ID NO: 2 is selected from ISIS NOs: 399841 or 399997.

In certain embodiments, a target region is nucleotides 6444-6463 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6444-6463 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 176. In certain such embodiments, an antisense compound targeted to nucleotides 6444-6463 of SEQ ID NO: 2 is selected from ISIS NOs: 410746.

In certain embodiments, a target region is nucleotides 6482-6518 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6482-6518 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 10, 177, 178, 179, 180 or 181. In certain such embodiments, an antisense compound targeted to nucleotides 6482-6518 of SEQ ID NO: 2 is selected from ISIS NOs: 395154, 399876, 406003, 406004, 406005, 406006 or 410747.

In certain embodiments, a target region is nucleotides 6492-6518 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6492-6518 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 10, 178, 179, 180 or 181. In certain such embodiments, an antisense compound targeted to nucleotides 6492-6518 of SEQ ID NO: 2 is selected from ISIS NOs: 395154, 399876, 406003, 406004, 406005 or 406006.

In certain embodiments, a target region is nucleotides 6528-6555 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6528-6555 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 182, 183, 184, 185 or 186. In certain such embodiments, an antisense compound targeted to nucleotides 6528-6555 of SEQ ID NO: 2 is selected from ISIS NOs: 406007, 410529, 410530, 410531, 410574, 410575, 410576, 410647, 410648, 410649 or 410748.

In certain embodiments, a target region is nucleotides 6528-6623 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6528-6623 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 13, 14, 15, 16, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, or 207. In certain such embodiments, an antisense compound targeted to nucleotides 6528-6623 of SEQ ID NO: 2 is selected from ISIS NOs: 395155, 395156, 395157, 399794, 399795, 399796, 399877, 399878, 399879, 399950, 399951, 399952, 406007, 406008, 406009, 406010, 406011, 406012, 406013, 406014, 406015, 406016, 406017, 406018, 406019, 406020, 410529, 410530, 410531, 410532, 410533, 410534, 410535, 410574, 410575, 410576, 410577, 410578, 410579, 410580, 410581, 410582, 410647, 410648, 410649, 410650, 410651, 410652, 410653, 410654, 410655, 410656, 410657, 410730, 410731, 410748, 410749, or 410750.

In certain embodiments, a target region is nucleotides 6534-6561 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6534-6561 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 184, 185, 186, 187, 188, 189, 190 or 191. In certain such embodiments, an antisense compound targeted to nucleotides 6534-6561 of SEQ ID NO: 2 is selected from ISIS NOs: 395155, 399877, 406008, 406009, 410529, 410530, 410531, 410532, 410533, 410534, 410574, 410575, 410576, 410577, 410578, 410579, 410580, 410581, 410647, 410648, 410649, 410650, 410651, 410652, 410653, 410654, or 410655.

In certain embodiments, a target region is nucleotides 6535-6562 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6535-6562 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 185, 186, 187, 188, 189, 190 or 191. In certain such embodiments, an antisense compound targeted to nucleotides 6535-6562 of SEQ ID NO: 2 is selected from ISIS NOs: 395155, 399794, 399877, 399950, 406008, 406009, 410530, 410531, 410532, 410533, 410534, 410575, 410576, 410577, 410578, 410579, 410580, 410581, 410648, 410649, 410650, 410651, 410652, 410653, 410654, 410655 or 410656.

In certain embodiments, a target region is nucleotides 6536-6563 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6536-6563 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 186, 187, 188, 189, 190, 191 or 192. In certain such embodiments, an antisense compound targeted to nucleotides 6536-6563 of SEQ ID NO: 2 is selected from ISIS NOs: 395155, 399794, 399877, 399950, 406008, 406009, 410531, 410532, 410533, 410534, 410535, 410576, 410577, 410578, 410579, 410580, 410581, 410582, 410649, 410650, 410651, 410652, 410653, 410654, 410655, 410656 or 410657.

In certain embodiments, a target region is nucleotides 6537-6563 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6537-6563 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 187, 188, 189, 190, 191 or 192. In certain such embodiments, an antisense compound targeted to nucleotides 6537-6563 of SEQ ID NO: 2 is selected from ISIS NOs: 395155, 399794, 399877, 399950, 406008, 406009, 410532, 410533, 410534, 410535, 410577, 410578, 410579, 410580, 410581, 410582, 410650, 410651, 410652, 410653, 410654, 410655, 410656 or 410657.

In certain embodiments, a target region is nucleotides 6538-6565 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6538-6565 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 187, 188, 189, 190, 191, 192 or 193. In certain such embodiments, an antisense compound targeted to nucleotides 6538-6565 of SEQ ID NO: 2 is selected from ISIS NOs: 399794, 399950, 406008, 406009, 406010, 410532, 410533, 410534, 410535, 410577, 410578, 410579, 410580, 410581, 410582, 410651, 410652, 410653, 410654, 410655, 410656 or 410657.

In certain embodiments, a target region is nucleotides 6539-6565 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6539-6565 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 188, 189, 190, 191, 192 or 193. In certain such embodiments, an antisense compound targeted to nucleotides 6539-6565 of SEQ ID NO: 2 is selected from ISIS NOs: 399794, 399950, 406008, 406009, 406010, 410533, 410534, 410535, 410578, 410579, 410580, 410581, 410582, 410652, 410653, 410654, 410655, 410656 or 410657.

In certain embodiments, a target region is nucleotides 6540-6567 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6540-6567 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 189, 190, 191, 192, 193 or 194. In certain such embodiments, an antisense compound targeted to nucleotides 6540-6567 of SEQ ID NO: 2 is selected from ISIS NOs: 399794, 399950, 406009, 406010, 406011, 410533, 410534, 410535, 410579, 410580, 410581, 410582, 410653, 410654, 410655, 410656 or 410657.

In certain embodiments, a target region is nucleotides 6541-6567 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6541-6567 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 190, 191, 192, 193 or 194. In certain such embodiments, an antisense compound targeted to nucleotides 6541-6567 of SEQ ID NO: 2 is selected from ISIS NOs: 399794, 399950, 406009, 406010, 406011, 410534, 410535, 410580, 410581, 410582, 410654, 410655, 410656 or 410657.

In certain embodiments, a target region is nucleotides 6542-6569 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6542-6569 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 191, 192, 193, 194 or 195. In certain such embodiments, an antisense compound targeted to nucleotides 6542-6569 of SEQ ID NO: 2 is selected from ISIS NOs: 399794, 399950, 406010, 406011, 406012, 410534, 410535, 410581, 410582, 410655, 410656 or 410657.

In certain embodiments, a target region is nucleotides 6546-6573 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6546-6573 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 13, 193, 194, 195 or 196. In certain such embodiments, an antisense compound targeted to nucleotides 6546-6573 of SEQ ID NO: 2 is selected from ISIS NOs: 399795, 399951, 406010, 406011, 406012 or 406013.

In certain embodiments, a target region is nucleotides 6557-6584 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6557-6584 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 14, 197 or 198. In certain such embodiments, an antisense compound targeted to nucleotides 6557-6584 of SEQ ID NO: 2 is selected from ISIS NOs: 395156, 399878, 410749 or 410750.

In certain embodiments, a target region is nucleotides 6575-6602 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6575-6602 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 15 or 198. In certain such embodiments, an antisense compound targeted to nucleotides 6575-6602 of SEQ ID NO: 2 is selected from ISIS NOs: 395157, 399879 or 410750.

In certain embodiments, a target region is nucleotides 6585-6611 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6585-6611 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 16, 199, 200 or 201. In certain such embodiments, an antisense compound targeted to nucleotides 6585-6611 of SEQ ID NO: 2 is selected from ISIS NOs: 399796, 399952, 406014, 406015 or 406016.

In certain embodiments, a target region is nucleotides 6594-6621 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6594-6621 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 202, 203, 204, 205 or 206. In certain such embodiments, an antisense compound targeted to nucleotides 6594-6621 of SEQ ID NO: 2 is selected from ISIS NOs: 406017, 406018, 406019, 406020 or 410730.

In certain embodiments, a target region is nucleotides 6596-6623 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6596-6623 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 203, 204, 205, 206 or 207. In certain such embodiments, an antisense compound targeted to nucleotides 6596-6623 of SEQ ID NO: 2 is selected from ISIS NOs: 406017, 406018, 406019, 406020 or 410731.

In certain embodiments, a target region is nucleotides 6652-6671 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6652-6671 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 108. In certain such embodiments, an antisense compound targeted to nucleotides 6652-6671 of SEQ ID NO: 2 is selected from ISIS NOs: 399842 or 399998.

In certain embodiments, a target region is nucleotides 7099-7118 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 7099-7118 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 114. In certain such embodiments, an antisense compound targeted to nucleotides 7099-7118 of SEQ ID NO: 2 is selected from ISIS NOs: 399843 or 399999.

In certain embodiments, a target region is nucleotides 7556-7584 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 7556-7584 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 91 or 131. In certain such embodiments, an antisense compound targeted to nucleotides 7556-7584 of SEQ ID NO: 2 is selected from ISIS NOs: 399844, 399845, 400000 or 400001.

In certain embodiments, a target region is nucleotides 8836-8855 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 8836-8855 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 145. In certain such embodiments, an antisense compound targeted to nucleotides 8836-8855 of SEQ ID NO: 2 is selected from ISIS NOs: 399846 or 400002.

In certain embodiments, a target region is nucleotides 8948-8967 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 8948-8967 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 90. In certain such embodiments, an antisense compound targeted to nucleotides 8948-8967 of SEQ ID NO: 2 is selected from ISIS NOs: 399847 or 400003.

In certain embodiments, a target region is nucleotides 9099-9118 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 9099-9118 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 125. In certain such embodiments, an antisense compound targeted to nucleotides 9099-9118 of SEQ ID NO: 2 is selected from ISIS NOs: 399848 or 400004.

In certain embodiments, a target region is nucleotides 9099-9168 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 9099-9168 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 17, 125 or 209. In certain such embodiments, an antisense compound targeted to nucleotides 9099-9168 of SEQ ID NO: 2 is selected from ISIS NOs: 395158, 399848, 399880, 400004 or 410752.

In certain embodiments, a target region is nucleotides 9130-9168 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 9130-9168 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 17. In certain such embodiments, an antisense compound targeted to nucleotides 9130-9168 of SEQ ID NO: 2 is selected from ISIS NOs: 395158 or 399880.

In certain embodiments, a target region is nucleotides 9207-9233 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 9207-9233 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 18, 210, 211, 212 or 213. In certain such embodiments, an antisense compound targeted to nucleotides 9207-9233 of SEQ ID NO: 2 is selected from ISIS NOs: 395159, 399881, 406021, 406022, 406023 or 406024.

In certain embodiments, a target region is nucleotides 9207-9235 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 9207-9235 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 18, 210, 211, 212, 213 or 214. In certain such embodiments, an antisense compound targeted to nucleotides 9207-9235 of SEQ ID NO: 2 is selected from ISIS NOs: 395159, 399881, 406021, 406022, 406023, 406024 or 410732.

In certain embodiments, a target region is nucleotides 9209-9235 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 9209-9235 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 18, 211, 212, 213 or 214. In certain such embodiments, an antisense compound targeted to nucleotides 9209-9235 of SEQ ID NO: 2 is selected from ISIS NOs: 395159, 399881, 406022, 406023, 406024 or 410732.

In certain embodiments, a target region is nucleotides 10252-10271 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 10252-10271 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 148. In certain such embodiments, an antisense compound targeted to nucleotides 10252-10271 of SEQ ID NO: 2 is selected from ISIS NOs: 399849 or 400005.

In certain embodiments, a target region is nucleotides 10633-10652 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 10633-10652 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 127. In certain such embodiments, an antisense compound targeted to nucleotides 10633-10652 of SEQ ID NO: 2 is selected from ISIS NOs: 395222 or 399944.

In certain embodiments, a target region is nucleotides 11308-11491 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 11308-11491 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 79 or 84. In certain such embodiments, an antisense compound targeted to nucleotides 11308-11491 of SEQ ID NO: 2 is selected from ISIS NOs: 395223, 399850, 399945 or 400006.

In certain embodiments, a target region is nucleotides 12715-12734 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 12715-12734 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 88. In certain such embodiments, an antisense compound targeted to nucleotides 12715-12734 of SEQ ID NO: 2 is selected from ISIS NOs: 399851 or 400007.

In certain embodiments, a target region is nucleotides 12928-12947 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 12928-12947 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 111. In certain such embodiments, an antisense compound targeted to nucleotides 12928-12947 of SEQ ID NO: 2 is selected from ISIS NOs: 399852 or 400008.

In certain embodiments, a target region is nucleotides 13681-13700 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 13681-13700 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 85. In certain such embodiments, an antisense compound targeted to nucleotides 13681-13700 of SEQ ID NO: 2 is selected from ISIS NOs: 395201 or 399923.

In certain embodiments, a target region is nucleotides 13746-13779 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 13746-13779 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 100 or 116. In certain such embodiments, an antisense compound targeted to nucleotides 13746-13779 of SEQ ID NO: 2 is selected from ISIS NOs: 399827, 399828, 399983 or 399984.

In certain embodiments, a target region is nucleotides 13816-13847 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 13816-13847 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 117 or 129. In certain such embodiments, an antisense compound targeted to nucleotides 13816-13847 of SEQ ID NO: 2 is selected from ISIS NOs: 395202, 399829, 399924 or 399985.

In certain embodiments, a target region is nucleotides 13903-13945 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 13903-13945 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 81 or 110. In certain such embodiments, an antisense compound targeted to nucleotides 13903-13945 of SEQ ID NO: 2 is selected from ISIS NOs: 395203, 399830, 399925 or 399986.

In certain embodiments, a target region is nucleotides 13977-14141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 13977-14141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 83, 136, 137, 140 or 152. In certain such embodiments, an antisense compound targeted to nucleotides 13977-14141 of SEQ ID NO: 2 is selected from ISIS NOs: 395204, 395205, 395206, 399831, 399832, 399926, 399927, 399928, 399987 or 399988.

In certain embodiments, a target region is nucleotides 14179-14198 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14179-14198 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 132. In certain such embodiments, an antisense compound targeted to nucleotides 14179-14198 of SEQ ID NO: 2 is selected from ISIS NOs: 395207 or 399929.

In certain embodiments, a target region is nucleotides 14267-14286 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14267-14286 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 139. In certain such embodiments, an antisense compound targeted to nucleotides 14267-14286 of SEQ ID NO: 2 is selected from ISIS NOs: 395208 or 399930.

In certain embodiments, a target region is nucleotides 14397-14423 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14397-14423 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 92 or 142. In certain such embodiments, an antisense compound targeted to nucleotides 14397-14423 of SEQ ID NO: 2 is selected from ISIS NOs: 395209, 399833, 399931 or 399989.

In certain embodiments, a target region is nucleotides 14441-14460 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14441-14460 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 113. In certain such embodiments, an antisense compound targeted to nucleotides 14441-14460 of SEQ ID NO: 2 is selected from ISIS NOs: 395210 or 399932.

In certain embodiments, a target region is nucleotides 14494-14513 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14494-14513 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 138. In certain such embodiments, an antisense compound targeted to nucleotides 14494-14513 of SEQ ID NO: 2 is selected from ISIS NOs: 395211 or 399933.

In certain embodiments, a target region is nucleotides 14494-14543 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14494-14543 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 98 or 138. In certain such embodiments, an antisense compound targeted to nucleotides 14494-14543 of SEQ ID NO: 2 is selected from ISIS NOs: 395211, 395212, 399933 or 399934.

In certain embodiments, a target region is nucleotides 14524-14543 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14524-14543 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 98. In certain such embodiments, an antisense compound targeted to nucleotides 14524-14543 of SEQ ID NO: 2 is selected from ISIS NOs: 395212 or 399934.

In certain embodiments, a target region is nucleotides 14601-14650 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14601-14650 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 96 or 124. In certain such embodiments, an antisense compound targeted to nucleotides 14601-14650 of SEQ ID NO: 2 is selected from ISIS NOs: 395213, 395214, 399935 or 399936.

In certain embodiments, a target region is nucleotides 14670-14700 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14670-14700 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 82, 103 or 133. In certain such embodiments, an antisense compound targeted to nucleotides 14670-14700 of SEQ ID NO: 2 is selected from ISIS NOs: 395215, 395216, 399834, 399937, 399938 or 399990.

In certain embodiments, a target region is nucleotides 14675-14700 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14675-14700 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 103 or 133. In certain such embodiments, an antisense compound targeted to nucleotides 14675-14700 of SEQ ID NO: 2 is selected from ISIS NOs: 395215, 395216, 399937 or 399938.

In certain embodiments, a target region is nucleotides 14801-14828 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14801-14828 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 155 or 156. In certain such embodiments, an antisense compound targeted to nucleotides 14801-14828 of SEQ ID NO: 2 is selected from ISIS NOs: 395217, 399835, 399939 or 399991.

In certain embodiments, a target region is nucleotides 14877-14912 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14877-14912 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 215, 216 or 217. In certain such embodiments, an antisense compound targeted to nucleotides 14877-14912 of SEQ ID NO: 2 is selected from ISIS NOs: 395160, 399882, 406025, 406026 or 410753.

In certain embodiments, a target region is nucleotides 14877-14915 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14877-14915 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20, 215, 216 or 217. In certain such embodiments, an antisense compound targeted to nucleotides 14877-14915 of SEQ ID NO: 2 is selected from ISIS NOs: 395160, 399797, 399882, 399953, 406025, 406026 or 410753.

In certain embodiments, a target region is nucleotides 14877-14973 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14877-14973 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20, 21, 22, 23, 24, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, or 233. In certain such embodiments, an antisense compound targeted to nucleotides 14877-14973 of SEQ ID NO: 2 is selected from ISIS NOs: 395160, 395161, 395162, 399797, 399798, 399799, 399882, 399883, 399884, 399953, 399954, 399955, 405869, 405870, 405871, 405872, 405873, 405874, 405875, 405876, 406025, 406026, 406027, 406028, 406029, 406030, 406031, 406032, 410733, 410753, or 410754.

In certain embodiments, a target region is nucleotides 14916-14943 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14916-14943 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 218, 219, 220, 221, 222 or 223. In certain such embodiments, an antisense compound targeted to nucleotides 14916-14943 of SEQ ID NO: 2 is selected from ISIS NOs: 395161, 399798, 399883, 399954, 405869, 405870, 405871, 405872, 405873 or 405874.

In certain embodiments, a target region is nucleotides 14916-14973 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14916-14973 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ IP NOs: 21, 22, 23, 24, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, or 223. In certain such embodiments, an antisense compound targeted to nucleotides 14916-14973 of SEQ ID NO: 2 is selected from ISIS NOs: 395161, 395162, 399798, 399799, 399883, 399884, 399954, 399955, 405869, 405870, 405871, 405872, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 406030, 406031, 406032, 410733, or 410754.

In certain embodiments, a target region is nucleotides 14925-14951 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14925-14951 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 224, 225, 226 or 227. In certain such embodiments, an antisense compound targeted to nucleotides 14925-14951 of SEQ ID NO: 2 is selected from ISIS NOs: 405875, 405876, 406027 or 406028.

In certain embodiments, a target region is nucleotides 14934-14963 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14934-14963 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 23, 228, 229, 230, 231 or 232. In certain such embodiments, an antisense compound targeted to nucleotides 14934-14963 of SEQ ID NO: 2 is selected from ISIS NOs: 399799, 399955, 406029, 406030, 406031, 406032 or 410733.

In certain embodiments, a target region is nucleotides 14946-14973 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14946-14973 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 24 or 233. In certain such embodiments, an antisense compound targeted to nucleotides 14946-14973 of SEQ ID NO: 2 is selected from ISIS NOs: 395162, 399884 or 410754.

In certain embodiments, a target region is nucleotides 14979-14998 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14979-14998 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 25. In certain such embodiments, an antisense compound targeted to nucleotides 14979-14998 of SEQ ID NO: 2 is selected from ISIS NOs: 395163 or 399885.

In certain embodiments, a target region is nucleotides 15254-15280 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15254-15280 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 235, 236 or 237. In certain such embodiments, an antisense compound targeted to nucleotides 15254-15280 of SEQ ID NO: 2 is selected from ISIS NOs: 405526, 405604, 406033 or 410756.

In certain embodiments, a target region is nucleotides 15254-15328 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15254-15328 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 235, 236, 237, 238, 239, 240, 241, 242, 243, 243, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15254-15328 of SEQ ID NO: 2 is selected from ISIS NOs: 395164, 395165, 399800, 399886, 399887, 399956, 405526, 405604, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 406033, 406034, 406035, 406036, 406037, 406038, 409126, 410536, 410537, 410538, 410539, 410583, 410584, 410585, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410593, 410594, 410595, 410658, 410659, 410660, 410661, 410662, 410663, 410664, 410665, 410666, 410667, 410668, 410669, 410670, 410671, 410756, 410757, or 410758.

In certain embodiments, a target region is nucleotides 15264-15290 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15264-15290 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 238 or 239. In certain such embodiments, an antisense compound targeted to nucleotides 15264-15290 of SEQ ID NO: 2 is selected from ISIS NOs: 395164, 399800, 399886, 399956, 406034, or 406035.

In certain embodiments, a target region is nucleotides 15279-15305 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15279-15305 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 240, 241 or 242. In certain such embodiments, an antisense compound targeted to nucleotides 15279-15305 of SEQ ID NO: 2 is selected from ISIS NOs: 406036, 406037, or 410757.

In certain embodiments, a target region is nucleotides 15291-15318 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15291-15318 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 243, 244, 245, 246, 247, 248, 249, 250, or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15291-15318 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881, 406038, 409126, 410536, 410537, 410583, 410584, 410585, 410586, 410587, 410588, 410589, 410590, 410658, 410659, 410660, 410661, 410662, 410663, 410664, 410665, or 410666.

In certain embodiments, a target region is nucleotides 15292-15319 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15292-15319 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 244, 245, 246, 247, 248, 249, 250 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15292-15319 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881, 406038, 409126, 410537, 410584, 410585, 410586, 410587, 410588, 410589, 410590, 410659, 410660, 410661, 410662, 410663, 410664, 410665, or 410666.

In certain embodiments, a target region is nucleotides 15293-15320 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15293-15320 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 245, 246, 247, 248, 249, 250, 251, 252 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15293-15320 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 406038, 409126, 410585, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410660, 410661, 410662, 410663, 410664, 410665, 410666, 410667, or 410668.

In certain embodiments, a target region is nucleotides 15294-15321 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15294-15321 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 246, 247, 248, 249, 250, 251, 252, 253 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15294-15321 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 409126, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410593, 410661, 410662, 410663, 410664, 410665, 410666, 410667, 410668 or 410669.

In certain embodiments, a target region is nucleotides 15294-15321 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15294-15321 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 246, 247, 248, 249, 250, 251, 252, 253 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15294-15321 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 409126, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410593, 410661, 410662, 410663, 410664, 410665, 410666, 410667, 410668 or 410669.

In certain embodiments, a target region is nucleotides 15295-15322 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15295-15322 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 247, 248, 249, 250, 250, 251, 252, 253, 254 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15295-15322 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 409126, 410538, 410587, 410588, 410589, 410590, 410591, 410592, 410593, 410594, 410662, 410663, 410664, 410665, 410666, 410667, 410668, 410669 or 410670.

In certain embodiments, a target region is nucleotides 15296-15315 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15296-15315 of SEQ ID NO: 2. In one such embodiment, an antisense compound targeted to nucleotides 15296-15315 of SEQ ID NO: 2 is ISIS NO: 405879.

In certain embodiments, a target region is nucleotides 15296-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15296-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 248, 249, 250, 251, 252, 253, 254, 255 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15296-15323 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405879, 405880, 405881, 405882, 405883, 405884, 409126, 410538, 410539, 410588, 410589, 410590, 410591, 410592, 410593, 410594, 410595, 410663, 410664, 410665, 410666, 410667, 410668, 410669, 410670 or 410671.

In certain embodiments, a target region is nucleotides 15297-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15297-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 249, 250, 251, 252, 253, 254, 255 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15297-15323 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405880, 405881, 405882, 405883, 405884, 409126, 410538, 410539, 410589, 410590, 410591, 410592, 410593, 410594, 410595, 410664, 410665, 410666, 410667, 410668, 410669, 410670 or 410671.

In certain embodiments, a target region is nucleotides 15298-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15298-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 250, 251, 252, 253, 254, 255 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15298-15323 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405881, 405882, 405883, 405884, 409126, 410538, 410539, 410590, 410591, 410592, 410593, 410594, 410595, 410665, 410666, 410667, 410668, 410669, 410670 or 410671.

In certain embodiments, a target region is nucleotides 15299-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15299-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 250, 251, 252, 253, 254 or 255. In certain such embodiments, an antisense compound targeted to nucleotides 15299-15323 of SEQ ID NO: 2 is selected from ISIS NOs: 405881, 405882, 405883, 405884, 410538, 410539, 410590, 410591, 410592, 410593, 410594, 410595, 410666, 410667, 410668, 410669, 410670 or 410671.

In certain embodiments, a target region is nucleotides 15300-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15300-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 251, 252, 253, 254 or 255. In certain such embodiments, an antisense compound targeted to nucleotides 15300-15323 of SEQ ID NO: 2 is selected from ISIS NOs: 405882, 405883, 405884, 410538, 410539, 410591, 410592, 410593, 410594, 410595, 410667, 410668, 410669, 410670 or 410671.

In certain embodiments, a target region is nucleotides 15301-15328 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15301-15328 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 252, 253, 254, 255 or 256. In certain such embodiments, an antisense compound targeted to nucleotides 15301-15328 of SEQ ID NO: 2 is selected from ISIS NOs: 405883, 405884, 410538, 410539, 410592, 410593, 410594, 410595, 410668, 410669, 410670, 410671 or 410758.

In certain embodiments, a target region is nucleotides 15330-15355 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15330-15355 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 29, 257, 258 or 259. In certain such embodiments, an antisense compound targeted to nucleotides 15330-15355 of SEQ ID NO: 2 is selected from ISIS NOs: 395166, 399888, 406039, 406040 or 406041.

In certain embodiments, a target region is nucleotides 15330-15490 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15330-15490 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 29, 30, 86, 257, 258, 259, 260, 261, 262 or 263. In certain such embodiments, an antisense compound targeted to nucleotides 15330-15490 of SEQ ID NO: 2 is selected from ISIS NOs: 395166, 399801, 399853, 399888, 399957, 400009, 406039, 406040, 406041, 406042, 406043, 406044 or 410759.

In certain embodiments, a target region is nucleotides 15339-15366 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15339-15366 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 30, 260, 261 or 262. In certain such embodiments, an antisense compound targeted to nucleotides 15339-15366 of SEQ ID NO: 2 is selected from ISIS NOs: 399801, 399957, 406042, 406043 or 406044.

In certain embodiments, a target region is nucleotides 15358-15490 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15358-15490 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 86 or 263. In certain such embodiments, an antisense compound targeted to nucleotides 15358-15490 of SEQ ID NO: 2 is selected from ISIS NOs: 399853, 400009 or 410759.

In certain embodiments, a target region is nucleotides 16134-16153 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 16134-16153 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 97. In certain such embodiments, an antisense compound targeted to nucleotides 16134-16153 of SEQ ID NO: 2 is selected from ISIS NOs: 399854 or 400010.

In certain embodiments, a target region is nucleotides 16668-16687 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 16668-16687 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 120. In certain such embodiments, an antisense compound targeted to nucleotides 16668-16687 of SEQ ID NO: 2 is selected from ISIS NOs: 399855 or 400011.

In certain embodiments, a target region is nucleotides 17267-17286 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 17267-17286 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 150. In certain such embodiments, an antisense compound targeted to nucleotides 17267-17286 of SEQ ID NO: 2 is selected from ISIS NOs: 399856 or 400012.

In certain embodiments, a target region is nucleotides 18377-18427 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18377-18427 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 115 or 134. In certain such embodiments, an antisense compound targeted to nucleotides 18377-18427 of SEQ ID NO: 2 is selected from ISIS NOs: 399857, 399858, 400013 or 400014.

In certain embodiments, a target region is nucleotides 18561-18580 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18561-18580 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 157. In certain such embodiments, an antisense compound targeted to nucleotides 18561-18580 of SEQ ID NO: 2 is selected from ISIS NOs: 395224 or 399946.

In certain embodiments, a target region is nucleotides 18591-18618 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18591-18618 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 32, 266, 267, 268 or 269. In certain such embodiments, an antisense compound targeted to nucleotides 18591-18618 of SEQ ID NO: 2 is selected from ISIS NOs: 395168, 399890, 405909, 405910, 405911 or 406045.

In certain embodiments, a target region is nucleotides 18591-18646 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18591-18646 of SEQ IL NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 32, 32, 266, 267, 268, 269, 270, 271 or 272. In certain such embodiments, an antisense compound targeted to nucleotides 18591-18646 of SEQ ID NO: 2 is selected from ISIS NOs: 395168, 399890, 405909, 405910, 405911, 405912, 406045, 410761 or 410762.

In certain embodiments, a target region is nucleotides 18591-18668 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18591-18668 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 32, 266, 267, 268, 269, 270, 271, 272 or 273. In certain such embodiments, an antisense compound targeted to nucleotides 18591-18668 of SEQ ID NO: 2 is selected from ISIS NOs: 395168, 399890, 405909, 405910, 405911, 405912, 406045, 410761, 410762 or 410763.

In certain embodiments, a target region is nucleotides 18695-18746 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18695-18746 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 33, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284 or 285. In certain such embodiments, an antisense compound targeted to nucleotides 18695-18746 of SEQ ID NO: 2 is selected from ISIS NOs: 395169, 399891, 405913, 405914, 405915, 405916, 405917, 405918, 405919, 405920, 405921, 405922, 410734 or 410764.

In certain embodiments, a target region is nucleotides 18705-18730 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18705-18730 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 33, 275, 276 or 277. In certain such embodiments, an antisense compound targeted to nucleotides 18705-18730 of SEQ ID NO: 2 is selected from ISIS NOs: 395169, 399891, 405913, 405914 or 405915.

In certain embodiments, a target region is nucleotides 18709-18736 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18709-18736 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 276, 277, 278, 279 or 280. In certain such embodiments, an antisense compound targeted to nucleotides 18709-18736 of SEQ ID NO: 2 is selected from ISIS NOs: 405914, 405915, 405916, 405917 or 410734.

In certain embodiments, a target region is nucleotides 18719-18746 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18719-18746 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 281, 282, 283, 284 or 285. In certain such embodiments, an antisense compound targeted to nucleotides 18719-18746 of SEQ ID NO: 2 is selected from ISIS NOs: 405918, 405919, 405920, 405921 or 405922.

In certain embodiments, a target region is nucleotides 19203-20080 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19203-20080 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 34, 35, 36, 37, 38, 39, 40, 41, 105, 128, 149, 151, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317 or 318. In certain such embodiments, an antisense compound targeted to nucleotides 19203-20080 of SEQ ID NO: 2 is selected from ISIS NOs: 395170, 395171, 395172, 395173, 395174, 395175, 399802, 399803, 399804, 399805, 399859, 399860, 399892, 399893, 399894, 399895, 399896, 399897, 399958, 399959, 399960, 399961, 400015, 400016, 405923, 405924, 405925, 405926, 405927, 405928, 405929, 405930, 405931, 405932, 405933, 405934, 405935, 405936, 405937, 405938, 405939, 405940, 405941, 405942, 405943, 405944, 405945, 405946, 405947, 410735, 410736, 410737, 410767, 410768 or 410769.

In certain embodiments, a target region is nucleotides 19931-19952 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19931-19952 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 149 or 188. In certain such embodiments, an antisense compound targeted to nucleotides 19931-19952 of SEQ ID NO: 2 is selected from ISIS NOs: 395170, 399892 or 405923.

In certain embodiments, a target region is nucleotides 19954-19981 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19954-19981 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 34, 290, 291, 292 or 293. In certain such embodiments, an antisense compound targeted to nucleotides 19954-19981 of SEQ ID NO: 2 is selected from ISIS NOs: 395171, 399893, 405924, 405925, 405926 or 410735.

In certain embodiments, a target region is nucleotides 19964-19990 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19964-19990 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 35, 128, 128, 294 or 295. In certain such embodiments, an antisense compound targeted to nucleotides 19964-19990 of SEQ ID NO: 2 is selected from ISIS NOs: 395172, 399802, 399894, 399958, 405927 or 405928.

In certain embodiments, a target region is nucleotides 19973-19999 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19973-19999 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 36, 296, 297 or 298. In certain such embodiments, an antisense compound targeted to nucleotides 19973-19999 of SEQ ID NO: 2 is selected from ISIS NOs: 395173, 399895, 405929, 405930 or 405931.

In certain embodiments, a target region is nucleotides 19982-20009 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19982-20009 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 37, 299, 300, 301 or 302. In certain such embodiments, an antisense compound targeted to nucleotides 19982-20009 of SEQ ID NO: 2 is selected from ISIS NOs: 399803, 399959, 405932, 405933, 405934 or 405935.

In certain embodiments, a target region is nucleotides 19992-20016 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19992-20016 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 38, 303 or 304. In certain such embodiments, an antisense compound targeted to nucleotides 19992-20016 of SEQ ID NO: 2 is selected from ISIS NOs: 395174, 399896, 405936 or 410736.

In certain embodiments, a target region is nucleotides 20016-20042 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20016-20042 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 305 or 306. In certain such embodiments, an antisense compound targeted to nucleotides 20016-20042 of SEQ ID NO: 2 is selected from ISIS NOs: 405937 or 410768.

In certain embodiments, a target region is nucleotides 20025-20052 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20025-20052 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 39, 307, 308, 309 or 310. In certain such embodiments, an antisense compound targeted to nucleotides 20025-20052 of SEQ ID NO: 2 is selected from ISIS NOs: 395175, 399897, 405938, 405939, 405940 or 405941.

In certain embodiments, a target region is nucleotides 20036-20062 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20036-20062 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 40, 311, 312, 313 or 314. In certain such embodiments, an antisense compound targeted to nucleotides 20036-20062 of SEQ ID NO: 2 is selected from ISIS NOs: 399804, 399960, 405942, 405943, 405944 or 410737.

In certain embodiments, a target region is nucleotides 20045-20070 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20045-20070 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 41, 315, 316 or 317. In certain such embodiments, an antisense compound targeted to nucleotides 20045-20070 of SEQ ID NO: 2 is selected from ISIS NOs: 399805, 399961, 405945, 405946 or 405947.

In certain embodiments, a target region is nucleotides 20100-20119 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20100-20119 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 158. In certain such embodiments, an antisense compound targeted to nucleotides 20100-20119 of SEQ ID NO: 2 is selected from ISIS NOs: 399861 or 400017.

In certain embodiments, a target region is nucleotides 20188-20207 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20188-20207 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 109. In certain such embodiments, an antisense compound targeted to nucleotides 20188-20207 of SEQ ID NO: 2 is selected from ISIS NOs: 399862 or 400018.

In certain embodiments, a target region is nucleotides 20624-20650 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20624-20650 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 141, 320 or 321. In certain such embodiments, an antisense compound targeted to nucleotides 20624-20650 of SEQ ID NO: 2 is selected from ISIS NOs: 399863, 400019, 405949 or 405950.

In certain embodiments, a target region is nucleotides 20624-20759 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20624-20759 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 44, 45, 46, 47, 48, 49, 50, 51, 87, 141, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358 or 359. In certain such embodiments, an antisense compound targeted to nucleotides 20624-20759 of SEQ ID NO: 2 is selected from ISIS NOs: 395177, 395178, 395179, 399807, 399808, 399809, 399810, 399811, 399812, 399813, 399863, 399899, 399900, 399901, 399963, 399964, 399965, 399966, 399967, 399968, 399969, 400019, 405557, 405885, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 405949, 405950, 405951, 405952, 405953, 405954, 405955, 405956, 405957, 405958, 405959, 405960, 405961, 405962, 405963, 405964, 405965, 405966, 405967, 405968, 405969, 405970, 405971, 405972, 405973, 405974, 408653, 410540, 410596, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681, 410682, 410738, 410739, 410740 or 410770.

In certain embodiments, a target region is nucleotides 20629-20804 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20629-20804 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 44, 45, 46, 47, 48, 49, 50, 51, 87, 119, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 356, 357, 358, 359 or 360. In certain such embodiments, an antisense compound targeted to nucleotides 20629-20804 of SEQ ID NO: 2 is selected from ISIS NOs: 395177, 395178, 395179, 395180, 399807, 399808, 399809, 399810, 399811, 399812, 399813, 399899, 399900, 399901, 399902, 399963, 399964, 399965, 399966, 399967, 399968, 399969, 405557, 405885, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 405949, 405950, 405951, 405952, 405953, 405954, 405955, 405956, 405957, 405958, 405959, 405960, 405961, 405962, 405963, 405964, 405965, 405966, 405967, 405968, 405969, 405970, 405971, 405972, 405973, 405974, 408653, 410540, 410596, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681, 410682, 410738, 410739, 410740, 410770 or 410771.

In certain embodiments, a target region is nucleotides 20633-20660 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20633-20660 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 322, 323, 324 or 325. In certain such embodiments, an antisense compound targeted to nucleotides 20633-20660 of SEQ ID NO: 2 is selected from ISIS NOs: 399807, 399963, 405951, 405952, 405953 or 410738.

In certain embodiments, a target region is nucleotides 20635-20781 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20635-20781 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 44, 45, 46, 47, 48, 49, 50, 51, 87, 119, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358 or 359. In certain such embodiments, an antisense compound targeted to nucleotides 20635-20781 of SEQ ID NO: 2 is selected from ISIS NOs: 395177, 395178, 395179, 395180, 399807, 399808, 399809, 399810, 399811, 399812, 399813, 399899, 399900, 399901, 399902, 399963, 399964, 399965, 399966, 399967, 399968, 399969, 405557, 405885, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 405952, 405953, 405954, 405955, 405956, 405957, 405958, 405959, 405960, 405961, 405962, 405963, 405964, 405965, 405966, 405967, 405968, 405969, 405970, 405971, 405972, 405973, 405974, 408653, 410540, 410596, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681, 410682, 410738, 410739, 410740 or 410770.

In certain embodiments, a target region is nucleotides 20643-20662 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20643-20662 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 326. In certain such embodiments, an antisense compound targeted to nucleotides 20643-20662 of SEQ ID NO: 2 is selected from ISIS NOs: 405954.

In certain embodiments, a target region is nucleotides 20657-20676 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20657-20676 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 327. In certain such embodiments, an antisense compound targeted to nucleotides 20657-20676 of SEQ ID NO: 2 is selected from ISIS NOs: 410770.

In certain embodiments, a target region is nucleotides 20670-20697 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20670-20697 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 328, 329, 330, 331 or 332. In certain such embodiments, an antisense compound targeted to nucleotides 20670-20697 of SEQ ID NO: 2 is selected from ISIS NOs: 405955, 405956, 405957, 405958 or 405959.

In certain embodiments, a target region is nucleotides 20680-20706 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20680-20706 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 333, 334, 335 or 336. In certain such embodiments, an antisense compound targeted to nucleotides 20680-20706 of SEQ ID NO: 2 is selected from ISIS NOs: 399808, 399964, 405960, 405961, 405962 or 410739.

In certain embodiments, a target region is nucleotides 20683-20781 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20683-20781 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 45, 46, 47, 48, 49, 50, 51, 87, 119, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358 or 359. In certain such embodiments, an antisense compound targeted to nucleotides 20683-20781 of SEQ ID NO: 2 is selected from ISIS NOs: 395177, 395178, 395179, 395180, 399808, 399809, 399810, 399811, 399812, 399813, 399899, 399900, 399901, 399902, 399964, 399965, 399966, 399967, 399968, 399969, 405557, 405885, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 405962, 405963, 405964, 405965, 405966, 405967, 405968, 405969, 405970, 405971, 405972, 405973, 405974, 408653, 410540, 410596, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681, 410682, 410739 or 410740.

In certain embodiments, a target region is nucleotides 20689-20715 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20689-20715 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 45, 46, 337 or 338. In certain such embodiments, an antisense compound targeted to nucleotides 20689-20715 of SEQ ID NO: 2 is selected from ISIS NOs: 395177, 399809, 399899, 399965, 405963 or 405964.

In certain embodiments, a target region is nucleotides 20698-20725 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20698-20725 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 47, 47, 339, 340, 341 or 342. In certain such embodiments, an antisense compound targeted to nucleotides 20698-20725 of SEQ ID NO: 2 is selected from ISIS NOs: 399810, 399966, 405965, 405966, 405967 or 405968.

In certain embodiments, a target region is nucleotides 20709-20736 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20709-20736 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 48, 48, 343, 344, 345 or 346. In certain such embodiments, an antisense compound targeted to nucleotides 20709-20736 of SEQ ID NO: 2 is selected from ISIS NOs: 399811, 399967, 405885, 405969, 405970 or 410740.

In certain embodiments, a target region is nucleotides 20717-20744 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20717-20744 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 346, 347, 348, 349, 350, 351, 352 or 353. In certain such embodiments, an antisense compound targeted to nucleotides 20717-20744 of SEQ ID NO: 2 is selected from ISIS NOs: 399812, 399968, 405885, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 410596, 410597, 410598, 410599, 410600, 410601, 410672, 410673, 410674, 410675, 410676, 410677 or 410678.

In certain embodiments, a target region is nucleotides 20718-20745 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20718-20745 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 347, 348, 349, 350, 351, 352 or 353. In certain such embodiments, an antisense compound targeted to nucleotides 20718-20745 of SEQ ID NO: 2 is selected from ISIS NOs: 395178, 399812, 399900, 399968, 405557, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 410596, 410597, 410598, 410599, 410600, 410601, 410672, 410673, 410674, 410675, 410676, 410677, 410678 or 410679.

In certain embodiments, a target region is nucleotides 20719-20746 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20719-20746 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 348, 349, 350, 351, 352, 353 or 354. In certain such embodiments, an antisense compound targeted to nucleotides 20719-20746 of SEQ ID NO: 2 is selected from ISIS NOs: 395178, 399812, 399900, 399968, 405557, 405887, 405888, 405889, 405890, 405891, 405892, 408653, 410596, 410597, 410598, 410599, 410600, 410601, 410602, 410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679 or 410680.

In certain embodiments, a target region is nucleotides 20720-20747 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20720-20747 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 349, 350, 351, 352, 353, 354 or 355. In certain such embodiments, an antisense compound targeted to nucleotides 20720-20747 of SEQ ID NO: 2 is selected from ISIS NOs: 395178, 399812, 399900, 399968, 405557, 405888, 405889, 405890, 405891, 405892, 405971, 408653, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680 or 410681.

In certain embodiments, a target region is nucleotides 20721-20748 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20721-20748 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 350, 351, 352, 353, 354, 355 or 356. In certain such embodiments, an antisense compound targeted to nucleotides 20721-20748 of SEQ ID NO: 2 is selected from ISIS NOs: 395178, 399812, 399900, 399968, 405557, 405889, 405890, 405891, 405892, 405971, 408653, 410540, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681 or 410682.

In certain embodiments, a target region is nucleotides 20722-20749 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20722-20749 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 50, 350, 351, 352, 353, 354, 355, 356 or 357. In certain such embodiments, an antisense compound targeted to nucleotides 20722-20749 of SEQ ID NO: 2 is selected from ISIS NOs: 395178, 399900, 405557, 405889, 405890, 405891, 405892, 405971, 405972, 408653, 410540, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410675, 410676, 410677, 410678, 410679, 410680, 410681 or 410682.

In certain embodiments, a target region is nucleotides 20727-20752 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20727-20752 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 87, 354, 355, 356 or 357. In certain such embodiments, an antisense compound targeted to nucleotides 20727-20752 of SEQ ID NO: 2 is selected from ISIS NOs: 395179, 399901, 405971, 405972, 408653, 410540, 410602, 410603, 410604, 410680, 410681 or 410682.

In certain embodiments, a target region is nucleotides 20735-20759 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20735-20759 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 51, 358 or 359. In certain such embodiments, an antisense compound targeted to nucleotides 20735-20759 of SEQ ID NO: 2 is selected from ISIS NOs: 399813, 399969, 405973 or 405974.

In certain embodiments, a target region is nucleotides 20762-21014 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20762-21014 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 93, 119 or 360. In certain such embodiments, an antisense compound targeted to nucleotides 20762-21014 of SEQ ID NO: 2 is selected from ISIS NOs: 395180, 399864, 399902, 400020 or 410771.

In certain embodiments, a target region is nucleotides 20785-21014 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20785-21014 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 93 or 360. In certain such embodiments, an antisense compound targeted to nucleotides 20785-21014 of SEQ ID NO: 2 is selected from ISIS NOs: 399864, 400020 or 410771.

In certain embodiments, a target region is nucleotides 21082-21107 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21082-21107 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 95 or 361. In certain such embodiments, an antisense compound targeted to nucleotides 21082-21107 of SEQ ID NO: 2 is selected from ISIS NOs: 399865, 400021 or 405975.

In certain embodiments, a target region is nucleotides 21082-21152 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21082-21152 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 53, 54, 95, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370 or 371. In certain such embodiments, an antisense compound targeted to nucleotides 21082-21152 of SEQ ID NO: 2 is selected from ISIS NOs: 395182, 399814, 399865, 399904, 399970, 400021, 405975, 405976, 405977, 405978, 405979, 405980, 405981, 405982, 405983, 410741 or 410772.

In certain embodiments, a target region is nucleotides 21091-21114 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21091-21114 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 53, 362 or 363. In certain such embodiments, an antisense compound targeted to nucleotides 21091-21114 of SEQ ID NO: 2 is selected from ISIS NOs: 399814, 399970, 405976 or 405977.

In certain embodiments, a target region is nucleotides 21118-21144 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21118-21144 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 54, 365, 366 or 367. In certain such embodiments, an antisense compound targeted to nucleotides 21118-21144 of SEQ ID NO: 2 is selected from ISIS NOs: 395182, 399904, 405978, 405979 or 405980.

In certain embodiments, a target region is nucleotides 21127-21152 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21127-21152 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 368, 369, 370 or 371. In certain such embodiments, an antisense compound targeted to nucleotides 21127-21152 of SEQ ID NO: 2 is selected from ISIS NOs: 405981, 405982, 405983 or 410741.

In certain embodiments, a target region is nucleotides 21181-21209 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21181-21209 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 372, 373, 374 or 375. In certain such embodiments, an antisense compound targeted to nucleotides 21181-21209 of SEQ ID NO: 2 is selected from ISIS NOs: 399815, 399971, 405564, 405641, 405984, 405985, 405986 or 405987.

In certain embodiments, a target region is nucleotides 21181-21211 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21181-21211 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 56, 372, 373, 374 or 375. In certain such embodiments, an antisense compound targeted to nucleotides 21181-21211 of SEQ ID NO: 2 is selected from ISIS NOs: 399815, 399816, 399971, 399972, 405564, 405641, 405984, 405985, 405986 or 405987.

In certain embodiments, a target region is nucleotides 21183-21211 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21183-21211 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 56, 373, 374 or 375. In certain such embodiments, an antisense compound targeted to nucleotides 21183-21211 of SEQ ID NO: 2 is selected from ISIS NOs: 399815, 399816, 399971, 399972, 405564, 405641, 405985, 405986 or 405987.

In certain embodiments, a target region is nucleotides 21481-21500 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21481-21500 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 143. In certain such embodiments, an antisense compound targeted to nucleotides 21481-21500 of SEQ ID NO: 2 is selected from ISIS NOs: 399866 or 400022.

In certain embodiments, a target region is nucleotides 21589-21608 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21589-21608 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 89. In certain such embodiments, an antisense compound targeted to nucleotides 21589-21608 of SEQ ID NO: 2 is selected from ISIS NOs: 399867 or 400023.

In certain embodiments, a target region is nucleotides 21692-21719 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21692-21719 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 123, 448, 449, 450, 451, 452, 453, 454 or 455. In certain such embodiments, an antisense compound targeted to nucleotides 21692-21719 of SEQ ID NO: 2 is selected from ISIS NOs: 399868, 400024, 406478, 406479, 406480, 406481, 406482, 406483, 406484 or 406485.

In certain embodiments, a target region is nucleotides 22000-22227 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22000-22227 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 389, 390, 391, 392, 393, 394, 395 or 396. In certain such embodiments, an antisense compound targeted to nucleotides 22000-22227 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 410550, 410551, 410552, 410553, 410554, 410555, 410616, 410617, 410618, 410619, 410620, 410621, 410622, 410623, 410695, 410696, 410697, 410698, 410699, 410700, 410701, 410702 or 410703.

In certain embodiments, a target region is nucleotides 22096-22115 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22096-22115 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 57. In certain such embodiments, an antisense compound targeted to nucleotides 22096-22115 of SEQ ID NO: 2 is selected from ISIS NOs: 395183 or 399905.

In certain embodiments, a target region is nucleotides 22096-22223 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22096-22223 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 57, 58, 59, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391 or 392. In certain such embodiments, an antisense compound targeted to nucleotides 22096-22223 of SEQ ID NO: 2 is selected from ISIS NOs: 395183, 395184, 395185, 399905, 399906, 399907, 405988, 405989, 405990, 410541, 410542, 410543, 410544, 410545, 410546, 410547, 410548, 410549, 410550, 410551, 410552, 410553, 410605, 410606, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410614, 410615, 410616, 410617, 410618, 410619, 410683, 410684, 410685, 410686, 410687, 410688, 410689, 410690, 410691, 410692, 410693, 410694, 410695, 410696, 410697, 410698, 410699 or 410773.

In certain embodiments, a target region is nucleotides 22096-22311 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22096-22311 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 57, 58, 59, 126, 126, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 401, 401, 402, 402, 402, 403, 403, 403, 404, 404, 404, 405, 405, 405 or 406. In certain such embodiments, an antisense compound targeted to nucleotides 22096-22311 of SEQ ID NO: 2 is selected from ISIS NOs: 395183, 395184, 395185, 395225, 399905, 399906, 399907, 399947, 405988, 405989, 405990, 405991, 405992, 405993, 405994, 405995, 410541, 410542, 410543, 410544, 410545, 410546, 410547, 410548, 410549, 410550, 410551, 410552, 410553, 410554, 410555, 410556, 410557, 410558, 410559, 410560, 410561, 410605, 410606, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410614, 410615, 410616, 410617, 410618, 410619, 410620, 410621, 410622, 410623, 410624, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410632, 410683, 410684, 410685, 410686, 410687, 410688, 410689, 410690, 410691, 410692, 410693, 410694, 410695, 410696, 410697, 410698, 410699, 410700, 410701, 410702, 410703, 410704, 410705, 410706, 410707, 410708, 410709, 410710, 410711, 410712, 410773 or 410774.

In certain embodiments, a target region is nucleotides 22133-22160 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22133-22160 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 376, 377, 378, 379, 380, 381, 382, 383 or 384. In certain such embodiments, an antisense compound targeted to nucleotides 22133-22160 of SEQ ID NO: 2 is selected from ISIS NOs: 405988, 405989, 410541, 410542, 410543, 410544, 410545, 410546, 410547, 410605, 410606, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410683, 410684, 410685, 410686, 410687, 410688, 410689, 410690 or 410691.

In certain embodiments, a target region is nucleotides 22133-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22133-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385 or 386. In certain such embodiments, an antisense compound targeted to nucleotides 22133-22163 of SEQ ID NO: 2 is selected from ISIS NOs: 395184, 399906, 405988, 405989, 405990, 410541, 410542, 410543, 410544, 410545, 410546, 410547, 410548, 410605, 410606, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410614, 410683, 410684, 410685, 410686, 410687, 410688, 410689, 410690, 410691, 410692 or 410693.

In certain embodiments, a target region is nucleotides 22134-22161 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22134-22161 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 377, 378, 379, 380, 381, 382, 383 or 384. In certain such embodiments, an antisense compound targeted to nucleotides 22134-22161 of SEQ ID NO: 2 is selected from ISIS NOs: 395184, 399906, 405988, 405989, 410542, 410543, 410544, 410545, 410546, 410547, 410606, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410684, 410685, 410686, 410687, 410688, 410689, 410690, 410691 or 410692.

In certain embodiments, a target region is nucleotides 22135-22162 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22135-22162 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 378, 379, 380, 381, 382, 383, 384 or 385. In certain such embodiments, an antisense compound targeted to nucleotides 22135-22162 of SEQ ID NO: 2 is selected from ISIS NOs: 395184, 399906, 405988, 405989, 410543, 410544, 410545, 410546, 410547, 410548, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410614, 410685, 410686, 410687, 410688, 410689, 410690, 410691, 410692 or 410693.

In certain embodiments, a target region is nucleotides 22136-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22136-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 379, 380, 381, 382, 383, 384, 385 or 386. In certain such embodiments, an antisense compound targeted to nucleotides 22136-22163 of SEQ ID NO: 2 is selected from ISIS NOs: 395184, 399906, 405988, 405989, 405990, 410544, 410545, 410546, 410547, 410548, 410608, 410609, 410610, 410611, 410612, 410613, 410614, 410686, 410687, 410688, 410689, 410690, 410691, 410692 or 410693.

In certain embodiments, a target region is nucleotides 22137-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22137-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 380, 381, 382, 383, 384, 385 or 386. In certain such embodiments, an antisense compound targeted to nucleotides 22137-22163 of SEQ ID NO: 2 is selected from ISIS NOs: 395184, 399906, 405988, 405989, 405990, 410545, 410546, 410547, 410548, 410609, 410610, 410611, 410612, 410613, 410614, 410687, 410688, 410689, 410690, 410691, 410692 or 410693.

In certain embodiments, a target region is nucleotides 22138-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22138-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 381, 382, 383, 384, 385 or 386. In certain such embodiments, an antisense compound targeted to nucleotides 22138-22163 of SEQ ID NO: 2 is selected from ISIS NOs: 395184, 399906, 405988, 405989, 405990, 410546, 410547, 410548, 410610, 410611, 410612, 410613, 410614, 410688, 410689, 410690, 410691, 410692 or 410693.

In certain embodiments, a target region is nucleotides 22189-22239 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22189-22239 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405 or 406. In certain such embodiments, an antisense compound targeted to nucleotides 22189-22239 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 405993, 405994, 405995, 410549, 410550, 410551, 410552, 410553, 410554, 410555, 410556, 410557, 410558, 410559, 410560, 410561, 410615, 410616, 410617, 410618, 410619, 410620, 410621, 410622, 410623, 410624, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410632, 410694, 410695, 410696, 410697, 410698, 410699, 410700, 410701, 410702, 410703, 410704, 410705, 410706, 410707, 410708, 410709, 410710, 410711, 410712, 410773 or 410774.

In certain embodiments, a target region is nucleotides 22199-22226 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22199-22226 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 388, 389, 390, 391, 392, 393, 394 or 395. In certain such embodiments, an antisense compound targeted to nucleotides 22199-22226 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 410549, 410550, 410551, 410552, 410553, 410554, 410555, 410615, 410616, 410617, 410618, 410619, 410620, 410621, 410622, 410694, 410695, 410696, 410697, 410698, 410699, 410700, 410701 or 410702.

In certain embodiments, a target region is nucleotides 22199-22227 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22199-22227 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 388, 389, 390, 391, 392, 393, 394, 395 or 396. In certain such embodiments, an antisense compound targeted to nucleotides 22199-22227 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 410549, 410550, 410551, 410552, 410553, 410554, 410555, 410615, 410616, 410617, 410618, 410619, 410620, 410621, 410622, 410623, 410694, 410695, 410696, 410697, 410698, 410699, 410700, 410701, 410702 or 410703.

In certain embodiments, a target region is nucleotides 22200-22227 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22200-22227 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 389, 390, 391, 392, 393, 394, 395 or 396. In certain such embodiments, an antisense compound targeted to nucleotides 22200-22227 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 410550, 410551, 410552, 410553, 410554, 410555, 410616, 410617, 410618, 410619, 410620, 410621, 410622, 410623, 410695, 410696, 410697, 410698, 410699, 410700, 410701, 410702 or 410703.

In certain embodiments, a target region is nucleotides 22201-22228 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22201-22228 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 390, 391, 392, 393, 394, 395, 396 or 397. In certain such embodiments, an antisense compound targeted to nucleotides 22201-22228 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 410551, 410552, 410553, 410554, 410555, 410556, 410617, 410618, 410619, 410620, 410621, 410622, 410623, 410624, 410696, 410697, 410698, 410699, 410700, 410701, 410702, 410703 or 410704.

In certain embodiments, a target region is nucleotides 22202-22229 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22202-22229 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 391, 392, 393, 394, 395, 396, 397 or 398. In certain such embodiments, an antisense compound targeted to nucleotides 22202-22229 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 405993, 410552, 410553, 410554, 410555, 410556, 410618, 410619, 410620, 410621, 410622, 410623, 410624, 410625, 410697, 410698, 410699, 410700, 410701, 410702, 410703, 410704 or 410705.

In certain embodiments, a target region is nucleotides 22203-22230 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22203-22230 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 392, 393, 394, 395, 396, 397, 398 or 399. In certain such embodiments, an antisense compound targeted to nucleotides 22203-22230 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 405993, 410553, 410554, 410555, 410556, 410557, 410619, 410620, 410621, 410622, 410623, 410624, 410625, 410626, 410698, 410699, 410700, 410701, 410702, 410703, 410704, 410705 or 410706.

In certain embodiments, a target region is nucleotides 22204-22231 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22204-22231 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 393, 394, 395, 396, 397, 398, 399 or 400. In certain such embodiments, an antisense compound targeted to nucleotides 22204-22231 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 405993, 405994, 410554, 410555, 410556, 410557, 410620, 410621, 410622, 410623, 410624, 410625, 410626, 410627, 410699, 410700, 410701, 410702, 410703, 410704, 410705, 410706 or 410707.

In certain embodiments, a target region is nucleotides 22205-22232 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22205-22232 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 393, 394, 395, 396, 397, 398, 399, 400 or 401. In certain such embodiments, an antisense compound targeted to nucleotides 22205-22232 of SEQ ID NO: 2 is selected from ISIS NOs: 405991, 405992, 405993, 405994, 410554, 410555, 410556, 410557, 410558, 410620, 410621, 410622, 410623, 410624, 410625, 410626, 410627, 410628, 410700, 410701, 410702, 410703, 410704, 410705, 410706, 410707 or 410708.

In certain embodiments, a target region is nucleotides 22206-22233 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22206-22233 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 394, 395, 396, 397, 398, 399, 400, 401 or 402. In certain such embodiments, an antisense compound targeted to nucleotides 22206-22233 of SEQ ID NO: 2 is selected from ISIS NOs: 405991, 405992, 405993, 405994, 405995, 410555, 410556, 410557, 410558, 410621, 410622, 410623, 410624, 410625, 410626, 410627, 410628, 410629, 410701, 410702, 410703, 410704, 410705, 410706, 410707, 410708 or 410709.

In certain embodiments, a target region is nucleotides 22207-22234 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22207-22234 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 395, 396, 397, 398, 399, 400, 401, 402 or 403. In certain such embodiments, an antisense compound targeted to nucleotides 22207-22234 of SEQ ID NO: 2 is selected from ISIS NOs: 405992, 405993, 405994, 405995, 410555, 410556, 410557, 410558, 410559, 410622, 410623, 410624, 410625, 410626, 410627, 410628, 410629, 410630, 410702, 410703, 410704, 410705, 410706, 410707, 410708, 410709 or 410710.

In certain embodiments, a target region is nucleotides 22208-22235 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22208-22235 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 396, 397, 398, 399, 400, 401, 402, 403 or 404. In certain such embodiments, an antisense compound targeted to nucleotides 22208-22235 of SEQ ID NO: 2 is selected from ISIS NOs: 405992, 405993, 405994, 405995, 410556, 410557, 410558, 410559, 410560, 410623, 410624, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410703, 410704, 410705, 410706, 410707, 410708, 410709, 410710 or 410711.

In certain embodiments, a target region is nucleotides 22209-22236 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22209-22236 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 397, 398, 399, 400, 401, 402, 403, 404 or 405. In certain such embodiments, an antisense compound targeted to nucleotides 22209-22236 of SEQ ID NO: 2 is selected from ISIS NOs: 405993, 405994, 405995, 410556, 410557, 410558, 410559, 410560, 410561, 410624, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410632, 410704, 410705, 410706, 410707, 410708, 410709, 410710, 410711 or 410712.

In certain embodiments, a target region is nucleotides 22210-22236 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22210-22236 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 398, 399, 400, 401, 402, 403, 404 or 405. In certain such embodiments, an antisense compound targeted to nucleotides 22210-22236 of SEQ ID NO: 2 is selected from ISIS NOs: 405993, 405994, 405995, 410557, 410558, 410559, 410560, 410561, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410632, 410705, 410706, 410707, 410708, 410709, 410710, 410711 or 410712.

In certain embodiments, a target region is nucleotides 22210-22239 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22210-22239 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 398, 399, 400, 401, 402, 403, 404, 405 or 405. In certain such embodiments, an antisense compound targeted to nucleotides 22210-22239 of SEQ ID NO: 2 is selected from ISIS NOs: 405993, 405994, 405995, 410557, 410558, 410559, 410560, 410561, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410632, 410705, 410706, 410707, 410708, 410709, 410710, 410711, 410712 or 410774.

In certain embodiments, a target region is nucleotides 22211-22236 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22211-22236 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 399, 400, 401, 402, 403, 404 or 405. In certain such embodiments, an antisense compound targeted to nucleotides 22211-22236 of SEQ ID NO: 2 is selected from ISIS NOs: 405994, 405995, 410557, 410558, 410559, 410560, 410561, 410626, 410627, 410628, 410629, 410630, 410631, 410632, 410706, 410707, 410708, 410709, 410710, 410711 or 410712.

In certain embodiments, a target region is nucleotides 22212-22239 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22212-22239 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 400, 401, 402, 403, 404, 405 or 406. In certain such embodiments, an antisense compound targeted to nucleotides 22212-22239 of SEQ ID NO: 2 is selected from ISIS NOs: 405994, 405995, 410558, 410559, 410560, 410561, 410627, 410628, 410629, 410630, 410631, 410632, 410707, 410708, 410709, 410710, 410711, 410712 or 410774.

In certain embodiments, a target region is nucleotides 22292-22311 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22292-22311 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 126. In certain such embodiments, an antisense compound targeted to nucleotides 22292-22311 of SEQ ID NO: 2 is selected from ISIS NOs: 395225 or 399947.

In certain embodiments, a target region is nucleotides 23985-24054 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 23985-24054 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 408, 409 or 410. In certain such embodiments, an antisense compound targeted to nucleotides 23985-24054 of SEQ ID NO: 2 is selected from ISIS NOs: 410776, 410777 or 410778.

In certain embodiments, a target region is nucleotides 24035-24134 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 24035-24134 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 410, 411, 412, 413, 414, 415 or 416. In certain such embodiments, an antisense compound targeted to nucleotides 24035-24134 of SEQ ID NO: 2 is selected from ISIS NOs: 395186, 399817, 399908, 399973, 405996, 405997, 410562, 410563, 410564, 410633, 410634, 410635, 410636, 410713, 410714, 410715, 410716, 410717, 410718, 410778 or 410779.

In certain embodiments, a target region is nucleotides 24095-24121 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 24095-24121 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413, 414 or 415. In certain such embodiments, an antisense compound targeted to nucleotides 24095-24121 of SEQ ID NO: 2 is selected from ISIS NOs: 395186, 399817, 399908, 399973, 405996, 405997, 410562, 410563, 410564, 410633, 410634, 410635, 410636, 410713, 410714, 410715, 410716, 410717 or 410718.

In certain embodiments, a target region is nucleotides 24858-24877 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 24858-24877 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 147. In certain such embodiments, an antisense compound targeted to nucleotides 24858-24877 of SEQ ID NO: 2 is selected from ISIS NOs: 395226 or 399948.

In certain embodiments, a target region is nucleotides 24907-24926 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 24907-24926 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 118. In certain such embodiments, an antisense compound targeted to nucleotides 24907-24926 of SEQ ID NO: 2 is selected from ISIS NOs: 399869 or 400025.

In certain embodiments, a target region is nucleotides 25413-25432 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 25413-25432 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 114. In certain such embodiments, an antisense compound targeted to nucleotides 25413-25432 of SEQ ID NO: 2 is selected from ISIS NOs: 399870 or 400026.

In certain embodiments, a target region is nucleotides 25994-26013 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 25994-26013 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 418. In certain such embodiments, an antisense compound targeted to nucleotides 25994-26013 of SEQ ID NO: 2 is selected from ISIS NOs: 410781.

In certain embodiments, a target region is nucleotides 26112-26139 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26112-26139 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422, 423, 424, 425 or 426. In certain such embodiments, an antisense compound targeted to nucleotides 26112-26139 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410565, 410566, 410567, 410568, 410569, 410570, 410571, 410637, 410638, 410639, 410640, 410641, 410642, 410643, 410644, 410719, 410720, 410721, 410722, 410723, 410724, 410725, 410726 or 410727.

In certain embodiments, a target region is nucleotides 26112-26161 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26112-26161 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429 or 430. In certain such embodiments, an antisense compound targeted to nucleotides 26112-26161 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410565, 410566, 410567, 410568, 410569, 410570, 410571, 410572, 410573, 410637, 410638, 410639, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410719, 410720, 410721, 410722, 410723, 410724, 410725, 410726, 410727, 410728, 410729, 410782 or 410783.

In certain embodiments, a target region is nucleotides 26112-27303 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26112-27303 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 112, 122, 135, 153, 154, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445 or 446. In certain such embodiments, an antisense compound targeted to nucleotides 26112-27303 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 395188, 395189, 395190, 395191, 395192, 395193, 395194, 395195, 395196, 395197, 395198, 395199, 399818, 399819, 399820, 399821, 399822, 399823, 399824, 399825, 399909, 399910, 399911, 399912, 399913, 399914, 399915, 399916, 399917, 399918, 399919, 399920, 399921, 399974, 399975, 399976, 399977, 399978, 399979, 399980, 399981, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405900, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405908, 405998, 410565, 410566, 410567, 410568, 410569, 410570, 410571, 410572, 410573, 410637, 410638, 410639, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410719, 410720, 410721, 410722, 410723, 410724, 410725, 410726, 410727, 410728, 410729, 410782 or 410783.

In certain embodiments, a target region is nucleotides 26113-26140 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26113-26140 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 420, 421, 422, 423, 424, 425, 426 or 427. In certain such embodiments, an antisense compound targeted to nucleotides 26113-26140 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410566, 410567, 410568, 410569, 410570, 410571, 410572, 410638, 410639, 410640, 410641, 410642, 410643, 410644, 410645, 410720, 410721, 410722, 410723, 410724, 410725, 410726, 410727 or 410728.

In certain embodiments, a target region is nucleotides 26114-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26114-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 421, 422, 423, 424, 425, 426, 427 or 248. In certain such embodiments, an antisense compound targeted to nucleotides 26114-26141 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410567, 410568, 410569, 410570, 410571, 410572, 410573, 410639, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410721, 410722, 410723, 410724, 410725, 410726, 410727, 410728 or 410729.

In certain embodiments, a target region is nucleotides 26115-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26115-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 422, 423, 424, 425, 426, 427 or 248. In certain such embodiments, an antisense compound targeted to nucleotides 26115-26141 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410568, 410569, 410570, 410571, 410572, 410573, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410722, 410723, 410724, 410725, 410726, 410727, 410728 or 410729.

In certain embodiments, a target region is nucleotides 26116-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26116-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 423, 424, 425, 426, 427 or 428. In certain such embodiments, an antisense compound targeted to nucleotides 26116-26141 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410569, 410570, 410571, 410572, 410573, 410641, 410642, 410643, 410644, 410645, 410646, 410723, 410724, 410725, 410726, 410727 or 410728.

In certain embodiments, a target region is nucleotides 26117-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26117-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 424, 425, 426, 427 or 428. In certain such embodiments, an antisense compound targeted to nucleotides 26117-26141 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410570, 410571, 410572, 410573, 410642, 410643, 410644, 410645, 410646, 410724, 410725, 410726, 410727, 410728 or 410729.

In certain embodiments, a target region is nucleotides 26117-26475 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26117-26475 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 63, 64, 65, 66, 67, 122, 153, 154, 424, 425, 426, 427, 428, 429 or 430. In certain such embodiments, an antisense compound targeted to nucleotides 26117-26475 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 395188, 395189, 395190, 395191, 395192, 399818, 399819, 399820, 399909, 399910, 399911, 399912, 399913, 399914, 399974, 399975, 399976, 405998, 410570, 410571, 410572, 410573, 410642, 410643, 410644, 410645, 410646, 410724, 410725, 410726, 410727, 410728, 410729, 410782 or 410783.

In certain embodiments, a target region is nucleotides 26118-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26118-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 424, 425, 426, 427 or 428. In certain such embodiments, an antisense compound targeted to nucleotides 26118-26141 of SEQ ID NO: 2 is selected from ISIS NOs: 405998, 410570, 410571, 410572, 410573, 410642, 410643, 410644, 410645, 410646, 410725, 410726, 410727, 410728 or 410729.

In certain embodiments, a target region is nucleotides 26120-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26120-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 426, 427 or 428. In certain such embodiments, an antisense compound targeted to nucleotides 26120-26141 of SEQ ID NO: 2 is selected from ISIS NOs: 405998, 410572, 410573, 410644, 410645, 410646, 410727, 410728 or 410729.

In certain embodiments, a target region is nucleotides 26132-26151 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26132-26151 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 429. In certain such embodiments, an antisense compound targeted to nucleotides 26132-26151 of SEQ ID NO: 2 is selected from ISIS NOs: 410782.

In certain embodiments, a target region is nucleotides 26142-26161 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26142-26161 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 430. In certain such embodiments, an antisense compound targeted to nucleotides 26142-26161 of SEQ ID NO: 2 is selected from ISIS NOs: 410783.

In certain embodiments, a target region is nucleotides 26217-26241 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26217-26241 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 63 or 154. In certain such embodiments, an antisense compound targeted to nucleotides 26217-26241 of SEQ ID NO: 2 is selected from ISIS NOs: 395188, 399818 or 399910.

In certain embodiments, a target region is nucleotides 26311-26335 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26311-26335 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 64 or 65. In certain such embodiments, an antisense compound targeted to nucleotides 26311-26335 of SEQ ID NO: 2 is selected from ISIS NOs: 395189, 399819, 399911 or 399975.

In certain embodiments, a target region is nucleotides 26389-26432 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26389-26432 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 66, 67 or 122. In certain such embodiments, an antisense compound targeted to nucleotides 26389-26432 of SEQ ID NO: 2 is selected from ISIS NOs: 395190, 395191, 399820, 399912, 399913 or 399976.

In certain embodiments, a target region is nucleotides 26456-26576 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26456-26576 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 68 or 153. In certain such embodiments, an antisense compound targeted to nucleotides 26456-26576 of SEQ ID NO: 2 is selected from ISIS NOs: 395192, 395193, 399914 or 399915.

In certain embodiments, a target region is nucleotides 26635-26662 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26635-26662 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 69, 431, 432, 433, 434, 435, 436, 437 or 438. In certain such embodiments, an antisense compound targeted to nucleotides 26635-26662 of SEQ ID NO: 2 is selected from ISIS NOs: 395194, 399916, 405893, 405894, 405895, 405896, 405897, 405898, 405899 or 405900.

In certain embodiments, a target region is nucleotides 26707-26734 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26707-26734 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441, 442, 443 or 444. In certain such embodiments, an antisense compound targeted to nucleotides 26707-26734 of SEQ ID NO: 2 is selected from ISIS NOs: 395195, 399821, 399917, 399977, 405901, 405902, 405903, 405904, 405905 or 405906.

In certain embodiments, a target region is nucleotides 26707-26736 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26707-26736 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441, 442, 443, 444, 445 or 446. In certain such embodiments, an antisense compound targeted to nucleotides 26707-26736 of SEQ ID NO: 2 is selected from ISIS NOs: 395195, 399821, 399917, 399977, 405901, 405902, 405903, 405904, 405905, 405906, 405907 or 405908.

In certain embodiments, a target region is nucleotides 26790-26820 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26790-26820 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 72, 73 or 135. In certain such embodiments, an antisense compound targeted to nucleotides 26790-26820 of SEQ ID NO: 2 is selected from ISIS NOs: 395196, 399822, 399823, 399918, 399978 or 399979.

In certain embodiments, a target region is nucleotides 27034-27263 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 27034-27263 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 74, 75 or 112. In certain such embodiments, an antisense compound targeted to nucleotides 27034-27263 of SEQ ID NO: 2 is selected from ISIS NOs: 395197, 395198, 399824, 399919, 399920 or 399980.

In certain embodiments, a target region is nucleotides 27279-27303 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 27279-27303 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 76 or 77. In certain such embodiments, an antisense compound targeted to nucleotides 27279-27303 of SEQ ID NO: 2 is selected from ISIS NOs: 395199, 399825, 399921 or 399981.

In certain embodiments, a target region is nucleotides 27350-27376 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 27350-27376 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 78 or 99. In certain such embodiments, an antisense compound targeted to nucleotides 27350-27376 of SEQ ID NO: 2 is selected from ISIS NOs: 395200, 399826, 399922 or 399982.

In certain embodiments, antisense compounds target a PCSK9 nucleic acid having the sequence of GENBANK® Accession No. AK124635.1, first deposited with GENBANK® on Sep. 8, 2003, and incorporated herein as SEQ ID NO: 3. In certain such embodiments, an antisense oligonucleotide is targeted to SEQ ID NO: 3. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 3 is at least 90% complementary to SEQ ID NO: 1. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 3 is at least 95% complementary to SEQ ID NO: 3. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 3 is 100% complementary to SEQ ID NO: 3. In certain such embodiments, an antisense oligonucleotide comprises a nucleotide sequence selected from a nucleotide sequence set forth in Table 12.

TABLE 12 Nucleotide sequences targeted to AK124635.1 (SEQ ID NO: 3) 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ ID NO NO: 3 NO: 3 Sequences (5′-3′) 85 155 174 ACAAATTCCCAGACTCAGCA 100 220 239 ATCTCAGGACAGGTGAGCAA 116 234 253 GAGTAGAGATTCTCATCTCA 129 290 309 GTGCCATCTGAACAGCACCT 117 302 321 GAGTCTTCTGAAGTGCCATC 81 377 396 AAGCAGGGCCTCAGGTGGAA 110 400 419 CCTGGAACCCCTGCAGCCAG 152 451 470 TTCAGGCAGGTTGCTGCTAG 83 460 479 AAGGAAGACTTCAGGCAGGT 140 472 491 TCAGCCAGGCCAAAGGAAGA 137 586 605 TAGGGAGAGCTCACAGATGC 136 596 615 TAGGAGAAAGTAGGGAGAGC 132 653 672 TAAAAGCTGCAAGAGACTCA 139 741 760 TCAGAGAAAACAGTCACCGA 92 871 890 AGAGACAGGAAGCTGCAGCT 142 878 897 TCATTTTAGAGACAGGAAGC 113 915 934 GAATAACAGTGATGTCTGGC 138 968 987 TCACAGCTCACCGAGTCTGC 98 998 1017 AGTGTAAAATAAAGCCCCTA 96 1075 1094 AGGACCCAAGTCATCCTGCT 124 1105 1124 GGCCATCAGCTGGCAATGCT 82 1144 1163 AAGGAAAGGGAGGCCTAGAG 133 1149 1168 TAGACAAGGAAAGGGAGGCC 103 1155 1174 ATTTCATAGACAAGGAAAGG 155 1275 1294 CTTATAGTTAACACACAGAA 156 1283 1302 AAGTCAACCTTATAGTTAAC 146 1315 1334 TGACATTTGTGGGAGAGGAG 161 1322 1341 TCCAAGGTGACATTTGTGGG 215 1351 1370 ATACACCTCCACCAGGCTGC 216 1362 1381 GTGTCTAGGAGATACACCTC 19 1365 1384 CTGGTGTCTAGGAGATACAC 217 1367 1386 TGCTGGTGTCTAGGAGATAC 20 1370 1389 GTATGCTGGTGTCTAGGAGA 21 1390 1409 GATTTCCCGGTGGTCACTCT 218 1392 1411 TCGATTTCCCGGTGGTCACT 219 1393 1412 CTCGATTTCCCGGTGGTCAC 220 1394 1413 CCTCGATTTCCCGGTGGTCA 221 1395 1414 CCCTCGATTTCCCGGTGGTC 22 1396 1415 GCCCTCGATTTCCCGGTGGT 222 1397 1416 TGCCCTCGATTTCCCGGTGG 223 1398 1417 CTGCCCTCGATTTCCCGGTG 224 1399 1418 CCTGCCCTCGATTTCCCGGT 225 1400 1419 CCCTGCCCTCGATTTCCCGG 226 1404 1423 ATGACCCTGCCCTCGATTTC 227 1406 1425 CCATGACCCTGCCCTCGATT 228 1408 1427 GACCATGACCCTGCCCTCGA 23 1410 1429 GTGACCATGACCCTGCCCTC 229 1412 1431 CGGTGACCATGACCCTGCCC 230 1414 1433 GTCGGTGACCATGACCCTGC 231 1416 1435 AAGTCGGTGACCATGACCCT 232 1418 1437 CGAAGTCGGTGACCATGACC 24 1420 1439 CTCGAAGTCGGTGACCATGA 233 1428 1447 GGCACATTCTCGAAGTCGGT 25 1453 1472 GTGGAAGCGGGTCCCGTCCT 234 1463 1482 TGGCCTGTCTGTGGAAGCGG 235 1490 1509 GGTGGGTGCCATGACTGTCA 236 1493 1512 CCAGGTGGGTGCCATGACTG 237 1497 1516 CCTGCCAGGTGGGTGCCATG 26 1500 1519 ACCCCTGCCAGGTGGGTGCC 238 1502 1521 CCACCCCTGCCAGGTGGGTG 27 1505 1524 TGACCACCCCTGCCAGGTGG 239 1507 1526 GCTGACCACCCCTGCCAGGT 240 1515 1534 TCCCGGCCGCTGACCACCCC 241 1519 1538 GGCATCCCGGCCGCTGACCA 242 1522 1541 GCCGGCATCCCGGCCGCTGA 243 1527 1546 GCCACGCCGGCATCCCGGCC 244 1528 1547 GGCCACGCCGGCATCCCGGC 245 1529 1548 TGGCCACGCCGGCATCCCGG 246 1530 1549 TTGGCCACGCCGGCATCCCG 247 1531 1550 CTTGGCCACGCCGGCATCCC 248 1532 1551 CCTTGGCCACGCCGGCATCC 249 1533 1552 CCCTTGGCCACGCCGGCATC 28 1534 1553 ACCCTTGGCCACGCCGGCAT 447 1534 1553 ACCCTTGGTCACGCCGGCAT 250 1535 1554 CACCCTTGGCCACGCCGGCA 251 1536 1555 GCACCCTTGGCCACGCCGGC 252 1537 1556 GGCACCCTTGGCCACGCCGG 253 1538 1557 TGGCACCCTTGGCCACGCCG 254 1539 1558 CTGGCACCCTTGGCCACGCC 255 1540 1559 GCTGGCACCCTTGGCCACGC 256 1545 1564 CGCATGCTGGCACCCTTGGC 257 1566 1585 CAGTTGAGCACGCGCAGGCT 258 1568 1587 GGCAGTTGAGCACGCGCAGG 29 1570 1589 TTGGCAGTTGAGCACGCGCA 259 1572 1591 CCTTGGCAGTTGAGCACGCG 30 1575 1594 TTCCCTTGGCAGTTGAGCAC 260 1577 1596 CCTTCCCTTGGCAGTTGAGC 261 1581 1600 GTGCCCTTCCCTTGGCAGTT 262 1583 1602 CCGTGCCCTTCCCTTGGCAG 263 1594 1613 GGTGCCGCTAACCGTGCCCT 264 1606 1625 CAGGCCTATGAGGGTGCCGC 31 1607 1626 CCAGGCCTATGAGGGTGCCG 458 1609 1622 GCCTATGAGGGTGC 459 1614 1627 TCCAGGCCTATGAG 265 1618 1637 CCGAATAAACTCCAGGCCTA 266 1626 1645 TGGCTTTTCCGAATAAACTC 32 1628 1647 GCTGGCTTTTCCGAATAAAC 267 1630 1649 CAGCTGGCTTTTCCGAATAA 268 1632 1651 ACCAGCTGGCTTTTCCGAAT 269 1634 1653 GGACCAGCTGGCTTTTCCGA 270 1638 1657 GGCTGGACCAGCTGGCTTTT 271 1649 1668 GTGGCCCCACAGGCTGGACC 272 1662 1681 AGCAGCACCACCAGTGGCCC 273 1684 1703 GCTGTACCCACCCGCCAGGG 274 1730 1749 CGACCCCAGCCCTCGCCAGG 33 1740 1759 GTGACCAGCACGACCCCAGC 275 1742 1761 CGGTGACCAGCACGACCCCA 276 1744 1763 AGCGGTGACCAGCACGACCC 277 1746 1765 GCAGCGGTGACCAGCACGAC 278 1748 1767 CGGCAGCGGTGACCAGCACG 279 1749 1768 CCGGCAGCGGTGACCAGCAC 280 1752 1771 TTGCCGGCAGCGGTGACCAG 281 1754 1773 AGTTGCCGGCAGCGGTGACC 282 1756 1775 GAAGTTGCCGGCAGCGGTGA 283 1758 1777 CGGAAGTTGCCGGCAGCGGT 284 1760 1779 CCCGGAAGTTGCCGGCAGCG 285 1762 1781 GTCCCGGAAGTTGCCGGCAG 306 1820 1839 GAGGCACCAATGATGTCCTC 39 1822 1841 TGGAGGCACCAATGATGTCC 307 1824 1843 GCTGGAGGCACCAATGATGT 308 1826 1845 TCGCTGGAGGCACCAATGAT 309 1828 1847 AGTCGCTGGAGGCACCAATG 310 1830 1849 GCAGTCGCTGGAGGCACCAA 40 1833 1852 GCTGCAGTCGCTGGAGGCAC 311 1835 1854 GTGCTGCAGTCGCTGGAGGC 312 1837 1856 AGGTGCTGCAGTCGCTGGAG 313 1839 1858 GCAGGTGCTGCAGTCGCTGG 314 1840 1859 AGCAGGTGCTGCAGTCGCTG 315 1842 1861 AAAGCAGGTGCTGCAGTCGC 41 1844 1863 ACAAAGCAGGTGCTGCAGTC 316 1846 1865 ACACAAAGCAGGTGCTGCAG 317 1848 1867 TGACACAAAGCAGGTGCTGC 318 1858 1877 TCCCACTCTGTGACACAAAG 101 1898 1917 ATGGCTGCAATGCCAGCCAC 319 1900 1919 TCATGGCTGCAATGCCAGCC 42 1903 1922 GCATCATGGCTGCAATGCCA 320 1905 1924 CAGCATCATGGCTGCAATGC 321 1907 1926 GACAGCATCATGGCTGCAAT 322 1909 1928 CAGACAGCATCATGGCTGCA 43 1911 1930 GGCAGACAGCATCATGGCTG 323 1913 1932 TCGGCAGACAGCATCATGGC 324 1915 1934 GCTCGGCAGACAGCATCATG 325 1917 1936 CGGCTCGGCAGACAGCATCA 326 1919 1938 TCCGGCTCGGCAGACAGCAT 327 1933 1952 CGGCCAGGGTGAGCTCCGGC 328 1946 1965 CTCTGCCTCAACTCGGCCAG 329 1948 1967 GTCTCTGCCTCAACTCGGCC 330 1950 1969 CAGTCTCTGCCTCAACTCGG 331 1952 1971 ATCAGTCTCTGCCTCAACTC 332 1954 1973 GGATCAGTCTCTGCCTCAAC 333 1956 1975 GTGGATCAGTCTCTGCCTCA 334 1958 1977 AAGTGGATCAGTCTCTGCCT 44 1959 1978 GAAGTGGATCAGTCTCTGCC 335 1961 1980 GAGAAGTGGATCAGTCTCTG 336 1963 1982 CAGAGAAGTGGATCAGTCTC 337 1965 1984 GGCAGAGAAGTGGATCAGTC 45 1967 1986 TTGGCAGAGAAGTGGATCAG 338 1969 1988 CTTTGGCAGAGAAGTGGATC 46 1972 1991 CATCTTTGGCAGAGAAGTGG 339 1974 1993 GACATCTTTGGCAGAGAAGT 340 1976 1995 ATGACATCTTTGGCAGAGAA 47 1978 1997 TGATGACATCTTTGGCAGAG 341 1980 1999 ATTGATGACATCTTTGGCAG 342 1982 2001 TCATTGATGACATCTTTGGC 48 1985 2004 GCCTCATTGATGACATCTTT 343 1987 2006 AGGCCTCATTGATGACATCT 344 1989 2008 CCAGGCCTCATTGATGACAT 345 1991 2010 AACCAGGCCTCATTGATGAC 346 1993 2012 GGAACCAGGCCTCATTGATG 347 1994 2013 GGGAACCAGGCCTCATTGAT 348 1995 2014 AGGGAACCAGGCCTCATTGA 349 1996 2615 CAGGGAACCAGGCCTCATTG 49 1997 2016 TCAGGGAACCAGGCCTCATT 350 1998 2017 CTCAGGGAACCAGGCCTCAT 351 1999 2018 CCTCAGGGAACCAGGCCTCA 352 2000 2019 TCCTCAGGGAACCAGGCCTC 353 2001 2020 GTCCTCAGGGAACCAGGCCT 50 2002 2021 GGTCCTCAGGGAACCAGGCC 354 2003 2022 TGGTCCTCAGGGAACCAGGC 355 2004 2023 CTGGTCCTCAGGGAACCAGG 356 2005 2024 GCTGGTCCTCAGGGAACCAG 357 2006 2025 CGCTGGTCCTCAGGGAACCA 87 2009 2028 ACCCGCTGGTCCTCAGGGAA 358 2011 2030 GTACCCGCTGGTCCTCAGGG 359 2013 2032 CAGTACCCGCTGGTCCTCAG 51 2016 2035 GGTCAGTACCCGCTGGTCCT 119 2038 2057 GCAGGGCGGCCACCAGGTTG 360 2061 2080 ACCTGCCCCATGGGTGCTGG 52 2073 2092 AAACAGCTGCCAACCTGCCC 361 2075 2094 CAAAACAGCTGCCAACCTGC 53 2078 2097 CTGCAAAACAGCTGCCAACC 362 2080 2099 TCCTGCAAAACAGCTGCCAA 363 2082 2101 AGTCCTGCAAAACAGCTGCC 104 2085 2104 CACAGTCCTGCAAAACAGCT 130 2095 2114 GTGCTGACCACACAGTCCTG 365 2105 2124 GGCCCCGAGTGTGCTGACCA 54 2108 2127 GTAGGCCCCGAGTGTGCTGA 366 2110 2129 GTGTAGGCCCCGAGTGTGCT 367 2112 2131 CCGTGTAGGCCCCGAGTGTG 368 2114 2133 ATCCGTGTAGGCCCCGAGTG 369 2116 2135 CCATCCGTGTAGGCCCCGAG 370 2118 2137 GGCCATCCGTGTAGGCCCCG 371 2120 2139 GTGGCCATCCGTGTAGGCCC 372 2168 2187 CTGGAGCAGCTCAGCAGCTC 460 2168 2181 CAGCTCAGCAGCTC 373 2170 2189 AACTGGAGCAGCTCAGCAGC 55 2173 2192 AGAAACTGGAGCAGCTCAGC 374 2175 2194 GGAGAAACTGGAGCAGCTCA 375 2177 2196 CTGGAGAAACTGGAGCAGCT 56 2179 2198 TCCTGGAGAAACTGGAGCAG 408 2245 2264 GTGCCAAGGTCCTCCACCTC 409 2265 2284 TCAGCACAGGCGGCTTGTGG 410 2295 2314 CCACGCACTGGTTGGGCTGA 60 2355 2374 CTTTGCATTCCAGACCTGGG 411 2356 2375 ACTTTGCATTCCAGACCTGG 412 2357 2376 GACTTTGCATTCCAGACCTG 413 2358 2377 TGACTTTGCATTCCAGACCT 414 2359 2378 TTGACTTTGCATTCCAGACC 61 2360 2379 CTTGACTTTGCATTCCAGAC 415 2362 2381 TCCTTGACTTTGCATTCCAG 416 2375 2394 CGGGATTCCATGCTCCTTGA 417 2405 2424 GCAGGCCACGGTCACCTGCT 418 2442 2461 GGAGGGCACTGCAGCCAGTC 419 2560 2579 CAGATGGCAACGGCTGTCAC 420 2561 2580 GCAGATGGCAACGGCTGTCA 421 2562 2581 AGCAGATGGCAACGGCTGTC 422 2563 2582 CAGCAGATGGCAACGGCTGT 423 2564 2583 GCAGCAGATGGCAACGGCTG 62 2565 2584 GGCAGCAGATGGCAACGGCT 424 2566 2585 CGGCAGCAGATGGCAACGGC 425 2567 2586 CCGGCAGCAGATGGCAACGG 426 2568 2587 TCCGGCAGCAGATGGCAACG 427 2569 2588 CTCCGGCAGCAGATGGCAAC 428 2570 2589 GCTCCGGCAGCAGATGGCAA 429 2580 2599 CCAGGTGCCGGCTCCGGCAG 430 2590 2609 GAGGCCTGCGCCAGGTGCCG 154 2665 2684 TTTTAAAGCTCAGCCCCAGC 63 2670 2689 AACCATTTTAAAGCTCAGCC 64 2759 2778 TCAAGGGCCAGGCCAGCAGC 65 2764 2783 CCCACTCAAGGGCCAGGCCA 122 2837 2856 GGAGGGAGCTTCCTGGCACC 66 2852 2871 ATGCCCCACAGTGAGGGAGG 67 2861 2880 AATGGTGAAATGCCCCACAG 153 2904 2923 TTGGGAGCAGCTGGCAGCAC 68 3005 3024 CATGGGAAGAATCCTGCCTC 431 3083 3102 ATGAGGGCCATCAGCACCTT 432 3084 3103 GATGAGGGCCATCAGCACCT 433 3085 3104 AGATGAGGGCCATCAGCACC 434 3086 3105 GAGATGAGGGCCATCAGCAC 69 3087 3106 GGAGATGAGGGCCATCAGCA 435 3088 3107 TGGAGATGAGGGCCATCAGC 436 3089 3108 CTGGAGATGAGGGCCATCAG 437 3090 3109 GCTGGAGATGAGGGCCATCA 438 3091 3110 AGCTGGAGATGAGGGCCATC 461 3132 3145 TTAATCAGGGAGCC 70 3155 3174 TAGATGCCATCCAGAAAGCT 439 3157 3176 GCTAGATGCCATCCAGAAAG 440 3158 3177 GGCTAGATGCCATCCAGAAA 441 3159 3178 TGGCTAGATGCCATCCAGAA 442 3160 3179 CTGGCTAGATGCCATCCAGA 71 3161 3180 TCTGGCTAGATGCCATCCAG 443 3162 3181 CTCTGGCTAGATGCCATCCA 444 3163 3182 CCTCTGGCTAGATGCCATCC 445 3164 3183 GCCTCTGGCTAGATGCCATC 446 3165 3184 AGCCTCTGGCTAGATGCCAT 72 3238 3257 GGCATAGAGCAGAGTAAAGG 73 3243 3262 AGCCTGGCATAGAGCAGAGT 135 3249 3268 TAGCACAGCCTGGCATAGAG 112 3482 3501 GAAGAGGCTTGGCTTCAGAG 74 3488 3507 AAGTAAGAAGAGGCTTGGCT 75 3692 3711 GCTCAAGGAGGGACAGTTGT 76 3727 3746 AAAGATAAATGTCTGCTTGC 77 3732 3751 ACCCAAAAGATAAATGTCTG 78 3798 3817 TCTTCAAGTTACAAAAGCAA 99 3805 3824 ATAAATATCTTCAAGTTACA

In certain embodiments, gapmer antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gapmer antisense compounds are targeted to SEQ ID NO: 3. In certain such embodiments, the nucleotide sequences illustrated in Table 12 have a 5-10-5 gapmer motif. Table 13 illustrates gapmer antisense compounds targeted to SEQ ID NO: 3, having a 5-10-5 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 13 Gapmer antisense compounds having a 5-10-5 motif targeted to SEQ ID NO: 3 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ ID Isis No Motif NO NO: 3 NO: 3 Mismatches Sequence (5′-3′) 395201 5-10-5 85 155 174 0 ACAAATTCCCAGACTCAGCA 399827 5-10-5 100 220 239 0 ATCTCAGGACAGGTGAGCAA 399828 5-10-5 116 234 253 0 GAGTAGAGATTCTCATCTCA 395202 5-10-5 129 290 309 0 GTGCCATCTGAACAGCACCT 399829 5-10-5 117 302 321 0 GAGTCTTCTGAAGTGCCATC 399830 5-10-5 81 377 396 0 AAGCAGGGCCTCAGGTGGAA 395203 5-10-5 110 400 419 0 CCTGGAACCCCTGCAGCCAG 395204 5-10-5 152 451 470 0 TTCAGGCAGGTTGCTGCTAG 399831 5-10-5 83 460 479 0 AAGGAAGACTTCAGGCAGGT 395205 5-10-5 140 472 491 0 TCAGCCAGGCCAAAGGAAGA 399832 5-10-5 137 586 605 0 TAGGGAGAGCTCACAGATGC 395206 5-10-5 136 596 615 0 TAGGAGAAAGTAGGGAGAGC 395207 5-10-5 132 653 672 0 TAAAAGCTGCAAGAGACTCA 395208 5-10-5 139 741 760 0 TCAGAGAAAACAGTCACCGA 399833 5-10-5 92 871 890 0 AGAGACAGGAAGCTGCAGCT 395209 5-10-5 142 878 897 0 TCATTTTAGAGACAGGAAGC 395210 5-10-5 113 915 934 0 GAATAACAGTGATGTCTGGC 395211 5-10-5 138 968 987 0 TCACAGCTCACCGAGTCTGC 395212 5-10-5 98 998 1017 0 AGTGTAAAATAAAGCCCCTA 395213 5-10-5 96 1075 1094 0 AGGACCCAAGTCATCCTGCT 395214 5-10-5 124 1105 1124 0 GGCCATCAGCTGGCAATGCT 399834 5-10-5 82 1144 1163 0 AAGGAAAGGGAGGCCTAGAG 395215 5-10-5 133 1149 1168 0 TAGACAAGGAAAGGGAGGCC 395216 5-10-5 103 1155 1174 0 ATTTCATAGACAAGGAAAGG 395217 5-10-5 155 1275 1294 0 CTTATAGTTAACACACAGAA 399835 5-10-5 156 1283 1302 0 AAGTCAACCTTATAGTTAAC 395218 5-10-5 146 1315 1334 0 TGACATTTGTGGGAGAGGAG 395219 5-10-5 161 1322 1341 0 TCCAAGGTGACATTTGTGGG 395160 5-10-5 19 1365 1384 0 CTGGTGTCTAGGAGATACAC 399797 5-10-5 20 1370 1389 0 GTATGCTGGTGTCTAGGAGA 395161 5-10-5 21 1390 1409 0 GATTTCCCGGTGGTCACTCT 399798 5-10-5 22 1396 1415 0 GCCCTCGATTTCCCGGTGGT 399799 5-10-5 23 1410 1429 0 GTGACCATGACCCTGCCCTC 395162 5-10-5 24 1420 1439 0 CTCGAAGTCGGTGACCATGA 395163 5-10-5 25 1453 1472 0 GTGGAAGCGGGTCCCGTCCT 395164 5-10-5 26 1500 1519 0 ACCCCTGCCAGGTGGGTGCC 399800 5-10-5 27 1505 1524 0 TGACCACCCCTGCCAGGTGG 395165 5-10-5 28 1534 1553 0 ACCCTTGGCCACGCCGGCAT 395166 5-10-5 29 1570 1589 0 TTGGCAGTTGAGCACGCGCA 399801 5-10-5 30 1575 1594 0 TTCCCTTGGCAGTTGAGCAC 395167 5-10-5 31 1607 1626 0 CCAGGCCTATGAGGGTGCCG 395168 5-10-5 32 1628 1647 0 GCTGGCTTTTCCGAATAAAC 395169 5-10-5 33 1740 1759 0 GTGACCAGCACGACCCCAGC 395175 5-10-5 39 1822 1841 0 TGGAGGCACCAATGATGTCC 399804 5-10-5 40 1833 1852 0 GCTGCAGTCGCTGGAGGCAC 399805 5-10-5 41 1844 1863 0 ACAAAGCAGGTGCTGCAGTC 395176 5-10-5 101 1898 1917 0 ATGGCTGCAATGCCAGCCAC 399806 5-10-5 42 1903 1922 0 GCATCATGGCTGCAATGCCA 399807 5-10-5 43 1911 1930 0 GGCAGACAGCATCATGGCTG 399808 5-10-5 44 1959 1978 0 GAAGTGGATCAGTCTCTGCC 395177 5-10-5 45 1967 1986 0 TTGGCAGAGAAGTGGATCAG 399809 5-10-5 46 1972 1991 0 CATCTTTGGCAGAGAAGTGG 399810 5-10-5 47 1978 1997 0 TGATGACATCTTTGGCAGAG 399811 5-10-5 48 1985 2004 0 GCCTCATTGATGACATCTTT 399812 5-10-5 49 1997 2016 0 TCAGGGAACCAGGCCTCATT 395178 5-10-5 50 2002 2021 0 GGTCCTCAGGGAACCAGGCC 395179 5-10-5 87 2009 2028 0 ACCCGCTGGTCCTCAGGGAA 399813 5-10-5 51 2016 2035 0 GGTCAGTACCCGCTGGTCCT 395180 5-10-5 119 2038 2057 0 GCAGGGCGGCCACCAGGTTG 395181 5-10-5 52 2073 2092 0 AAACAGCTGCCAACCTGCCC 399814 5-10-5 53 2078 2097 0 CTGCAAAACAGCTGCCAACC 395220 5-10-5 104 2085 2104 0 CACAGTCCTGCAAAACAGCT 399836 5-10-5 130 2095 2114 0 GTGCTGACCACACAGTCCTG 395182 5-10-5 54 2108 2127 0 GTAGGCCCCGAGTGTGCTGA 399815 5-10-5 55 2173 2192 0 AGAAACTGGAGCAGCTCAGC 399816 5-10-5 56 2179 2198 0 TCCTGGAGAAACTGGAGCAG 395186 5-10-5 60 2355 2374 0 CTTTGCATTCCAGACCTGGG 399817 5-10-5 61 2360 2379 0 CTTGACTTTGCATTCCAGAC 395187 5-10-5 62 2565 2584 0 GGCAGCAGATGGCAACGGCT 395188 5-10-5 154 2665 2684 0 TTTTAAAGCTCAGCCCCAGC 399818 5-10-5 63 2670 2689 0 AACCATTTTAAAGCTCAGCC 395189 5-10-5 64 2759 2778 0 TCAAGGGCCAGGCCAGCAGC 399819 5-10-5 65 2764 2783 0 CCCACTCAAGGGCCAGGCCA 399820 5-10-5 122 2837 2856 0 GGAGGGAGCTTCCTGGCACC 395190 5-10-5 66 2852 2871 0 ATGCCCCACAGTGAGGGAGG 395191 5-10-5 67 2861 2880 0 AATGGTGAAATGCCCCACAG 395192 5-10-5 153 2904 2923 0 TTGGGAGCAGCTGGCAGCAC 395193 5-10-5 68 3005 3024 0 CATGGGAAGAATCCTGCCTC 395194 5-10-5 69 3087 3106 0 GGAGATGAGGGCCATCAGCA 395195 5-10-5 70 3155 3174 0 TAGATGCCATCCAGAAAGCT 399821 5-10-5 71 3161 3180 0 TCTGGCTAGATGCCATCCAG 395196 5-10-5 72 3238 3257 0 GGCATAGAGCAGAGTAAAGG 399822 5-10-5 73 3243 3262 0 AGCCTGGCATAGAGCAGAGT 399823 5-10-5 135 3249 3268 0 TAGCACAGCCTGGCATAGAG 395197 5-10-5 112 3482 3501 0 GAAGAGGCTTGGCTTCAGAG 399824 5-10-5 74 3488 3507 0 AAGTAAGAAGAGGCTTGGCT 395198 5-10-5 75 3692 3711 0 GCTCAAGGAGGGACAGTTGT 395199 5-10-5 76 3727 3746 0 AAAGATAAATGTCTGCTTGC 399825 5-10-5 77 3732 3751 0 ACCCAAAAGATAAATGTCTG 395200 5-10-5 78 3798 3817 0 TCTTCAAGTTACAAAAGCAA 399826 5-10-5 99 3805 3824 0 ATAAATATCTTCAAGTTACA 405869 5-10-5 218 1392 1411 0 TCGATTTCCCGGTGGTCACT 405870 5-10-5 219 1393 1412 0 CTCGATTTCCCGGTGGTCAC 405871 5-10-5 220 1394 1413 0 CCTCGATTTCCCGGTGGTCA 405872 5-10-5 221 1395 1414 0 CCCTCGATTTCCCGGTGGTC 405873 5-10-5 222 1397 1416 0 TGCCCTCGATTTCCCGGTGG 405874 5-10-5 223 1398 1417 0 CTGCCCTCGATTTCCCGGTG 405875 5-10-5 224 1399 1418 0 CCTGCCCTCGATTTCCCGGT 405876 5-10-5 225 1400 1419 0 CCCTGCCCTCGATTTCCCGG 405877 5-10-5 246 1530 1549 0 TTGGCCACGCCGGCATCCCG 405878 5-10-5 247 1531 1550 0 CTTGGCCACGCCGGCATCCC 405879 5-10-5 248 1532 1551 0 CCTTGGCCACGCCGGCATCC 405880 5-10-5 249 1533 1552 0 CCCTTGGCCACGCCGGCATC 405881 5-10-5 250 1535 1554 0 CACCCTTGGCCACGCCGGCA 405882 5-10-5 251 1536 1555 0 GCACCCTTGGCCACGCCGGC 405883 5-10-5 252 1537 1556 0 GGCACCCTTGGCCACGCCGG 405884 5-10-5 253 1538 1557 0 TGGCACCCTTGGCCACGCCG 405885 5-10-5 346 1993 2012 0 GGAACCAGGCCTCATTGATG 405886 5-10-5 347 1994 2013 0 GGGAACCAGGCCTCATTGAT 405887 5-10-5 348 1995 2014 0 AGGGAACCAGGCCTCATTGA 405888 5-10-5 349 1996 2015 0 CAGGGAACCAGGCCTCATTG 405889 5-10-5 350 1998 2017 0 CTCAGGGAACCAGGCCTCAT 405890 5-10-5 351 1999 2018 0 CCTCAGGGAACCAGGCCTCA 405891 5-10-5 352 2000 2019 0 TCCTCAGGGAACCAGGCCTC 405892 5-10-5 353 2001 2020 0 GTCCTCAGGGAACCAGGCCT 405893 5-10-5 431 3083 3102 0 ATGAGGGCCATCAGCACCTT 405894 5-10-5 432 3084 3103 0 GATGAGGGCCATCAGCACCT 405895 5-10-5 433 3085 3104 0 AGATGAGGGCCATCAGCACC 405896 5-10-5 434 3086 3105 0 GAGATGAGGGCCATCAGCAC 405897 5-10-5 435 3088 3107 0 TGGAGATGAGGGCCATCAGC 405898 5-10-5 436 3089 3108 0 CTGGAGATGAGGGCCATCAG 405899 5-10-5 437 3090 3109 0 GCTGGAGATGAGGGCCATCA 405900 5-10-5 438 3091 3110 0 AGCTGGAGATGAGGGCCATC 405901 5-10-5 439 3157 3176 0 GCTAGATGCCATCCAGAAAG 405902 5-10-5 440 3158 3177 0 GGCTAGATGCCATCCAGAAA 405903 5-10-5 441 3159 3178 0 TGGCTAGATGCCATCCAGAA 405904 5-10-5 442 3160 3179 0 CTGGCTAGATGCCATCCAGA 405905 5-10-5 443 3162 3181 0 CTCTGGCTAGATGCCATCCA 405906 5-10-5 444 3163 3182 0 CCTCTGGCTAGATGCCATCC 405907 5-10-5 445 3164 3183 0 GCCTCTGGCTAGATGCCATC 405908 5-10-5 446 3165 3184 0 AGCCTCTGGCTAGATGCCAT 405909 5-10-5 267 1630 1649 0 CAGCTGGCTTTTCCGAATAA 405910 5-10-5 268 1632 1651 0 ACCAGCTGGCTTTTCCGAAT 405911 5-10-5 269 1634 1653 0 GGACCAGCTGGCTTTTCCGA 405912 5-10-5 270 1638 1657 0 GGCTGGACCAGCTGGCTTTT 405913 5-10-5 275 1742 1761 0 CGGTGACCAGCACGACCCCA 405914 5-10-5 276 1744 1763 0 AGCGGTGACCAGCACGACCC 405915 5-10-5 277 1746 1765 0 GCAGCGGTGACCAGCACGAC 405916 5-10-5 278 1748 1767 0 CGGCAGCGGTGACCAGCACG 405917 5-10-5 280 1752 1771 0 TTGCCGGCAGCGGTGACCAG 405918 5-10-5 281 1754 1773 0 AGTTGCCGGCAGCGGTGACC 405919 5-10-5 282 1756 1775 0 GAAGTTGCCGGCAGCGGTGA 405920 5-10-5 283 1758 1777 0 CGGAAGTTGCCGGCAGCGGT 405921 5-10-5 284 1760 1779 0 CCCGGAAGTTGCCGGCAGCG 405922 5-10-5 285 1762 1781 0 GTCCCGGAAGTTGCCGGCAG 405937 5-10-5 306 1820 1839 0 GAGGCACCAATGATGTCCTC 405938 5-10-5 307 1824 1843 0 GCTGGAGGCACCAATGATGT 405939 5-10-5 308 1826 1845 0 TCGCTGGAGGCACCAATGAT 405940 5-10-5 309 1828 1847 0 AGTCGCTGGAGGCACCAATG 405941 5-10-5 310 1830 1849 0 GCAGTCGCTGGAGGCACCAA 405942 5-10-5 311 1835 1854 0 GTGCTGCAGTCGCTGGAGGC 405943 5-10-5 312 1837 1856 0 AGGTGCTGCAGTCGCTGGAG 405944 5-10-5 314 1840 1859 0 AGCAGGTGCTGCAGTCGCTG 405945 5-10-5 315 1842 1861 0 AAAGCAGGTGCTGCAGTCGC 405946 5-10-5 316 1846 1865 0 ACACAAAGCAGGTGCTGCAG 405947 5-10-5 317 1848 1867 0 TGACACAAAGCAGGTGCTGC 405948 5-10-5 319 1900 1919 0 TCATGGCTGCAATGCCAGCC 405949 5-10-5 320 1905 1924 0 CAGCATCATGGCTGCAATGC 405950 5-10-5 321 1907 1926 0 GACAGCATCATGGCTGCAAT 405951 5-10-5 322 1909 1928 0 CAGACAGCATCATGGCTGCA 405952 5-10-5 323 1913 1932 0 TCGGCAGACAGCATCATGGC 405953 5-10-5 325 1917 1936 0 CGGCTCGGCAGACAGCATCA 405954 5-10-5 326 1919 1938 0 TCCGGCTCGGCAGACAGCAT 405955 5-10-5 328 1946 1965 0 CTCTGCCTCAACTCGGCCAG 405956 5-10-5 329 1948 1967 0 GTCTCTGCCTCAACTCGGCC 405957 5-10-5 330 1950 1969 0 CAGTCTCTGCCTCAACTCGG 405958 5-10-5 331 1952 1971 0 ATCAGTCTCTGCCTCAACTC 405959 5-10-5 332 1954 1973 0 GGATCAGTCTCTGCCTCAAC 405960 5-10-5 333 1956 1975 0 GTGGATCAGTCTCTGCCTCA 405961 5-10-5 334 1958 1977 0 AAGTGGATCAGTCTCTGCCT 405962 5-10-5 335 1961 1980 0 GAGAAGTGGATCAGTCTCTG 405963 5-10-5 337 1965 1984 0 GGCAGAGAAGTGGATCAGTC 405964 5-10-5 338 1969 1988 0 CTTTGGCAGAGAAGTGGATC 405965 5-10-5 339 1974 1993 0 GACATCTTTGGCAGAGAAGT 405966 5-10-5 340 1976 1995 0 ATGACATCTTTGGCAGAGAA 405967 5-10-5 341 1980 1999 0 ATTGATGACATCTTTGGCAG 405968 5-10-5 342 1982 2001 0 TCATTGATGACATCTTTGGC 405969 5-10-5 343 1987 2006 0 AGGCCTCATTGATGACATCT 405970 5-10-5 344 1989 2008 0 CCAGGCCTCATTGATGACAT 405971 5-10-5 355 2004 2023 0 CTGGTCCTCAGGGAACCAGG 405972 5-10-5 357 2006 2025 0 CGCTGGTCCTCAGGGAACCA 405973 5-10-5 358 2011 2030 0 GTACCCGCTGGTCCTCAGGG 405974 5-10-5 359 2013 2032 0 CAGTACCCGCTGGTCCTCAG 405975 5-10-5 361 2075 2094 0 CAAAACAGCTGCCAACCTGC 405976 5-10-5 362 2080 2099 0 TCCTGCAAAACAGCTGCCAA 405977 5-10-5 363 2082 2101 0 AGTCCTGCAAAACAGCTGCC 405978 5-10-5 365 2105 2124 0 GGCCCCGAGTGTGCTGACCA 405979 5-10-5 366 2110 2129 0 GTGTAGGCCCCGAGTGTGCT 405980 5-10-5 367 2112 2131 0 CCGTGTAGGCCCCGAGTGTG 405981 5-10-5 368 2114 2133 0 ATCCGTGTAGGCCCCGAGTG 405982 5-10-5 370 2118 2137 0 GGCCATCCGTGTAGGCCCCG 405983 5-10-5 371 2120 2139 0 GTGGCCATCCGTGTAGGCCC 405984 5-10-5 372 2168 2187 0 CTGGAGCAGCTCAGCAGCTC 405985 5-10-5 373 2170 2189 0 AACTGGAGCAGCTCAGCAGC 405986 5-10-5 374 2175 2194 0 GGAGAAACTGGAGCAGCTCA 405987 5-10-5 375 2177 2196 0 CTGGAGAAACTGGAGCAGCT 405996 5-10-5 412 2357 2376 0 GACTTTGCATTCCAGACCTG 405997 5-10-5 415 2362 2381 0 TCCTTGACTTTGCATTCCAG 405998 5-10-5 426 2568 2587 0 TCCGGCAGCAGATGGCAACG 406025 5-10-5 216 1362 1381 0 GTGTCTAGGAGATACACCTC 406026 5-10-5 217 1367 1386 0 TGCTGGTGTCTAGGAGATAC 406027 5-10-5 226 1404 1423 0 ATGACCCTGCCCTCGATTTC 406028 5-10-5 227 1406 1425 0 CCATGACCCTGCCCTCGATT 406029 5-10-5 228 1408 1427 0 GACCATGACCCTGCCCTCGA 406030 5-10-5 229 1412 1431 0 CGGTGACCATGACCCTGCCC 406031 5-10-5 230 1414 1433 0 GTCGGTGACCATGACCCTGC 406032 5-10-5 232 1418 1437 0 CGAAGTCGGTGACCATGACC 406033 5-10-5 237 1497 1516 0 CCTGCCAGGTGGGTGCCATG 406034 5-10-5 238 1502 1521 0 CCACCCCTGCCAGGTGGGTG 406035 5-10-5 239 1507 1526 0 GCTGACCACCCCTGCCAGGT 406036 5-10-5 241 1519 1538 0 GGCATCCCGGCCGCTGACCA 406037 5-10-5 242 1522 1541 0 GCCGGCATCCCGGCCGCTGA 406038 5-10-5 245 1529 1548 0 TGGCCACGCCGGCATCCCGG 406039 5-10-5 257 1566 1585 0 CAGTTGAGCACGCGCAGGCT 406040 5-10-5 258 1568 1587 0 GGCAGTTGAGCACGCGCAGG 406041 5-10-5 259 1572 1591 0 CCTTGGCAGTTGAGCACGCG 406042 5-10-5 260 1577 1596 0 CCTTCCCTTGGCAGTTGAGC 406043 5-10-5 261 1581 1600 0 GTGCCCTTCCCTTGGCAGTT 406044 5-10-5 262 1583 1602 0 CCGTGCCCTTCCCTTGGCAG 406045 5-10-5 266 1626 1645 0 TGGCTTTTCCGAATAAACTC 408642 5-10-5 264 1606 1625 0 CAGGCCTATGAGGGTGCCGC 408653 5-10-5 354 2003 2022 0 TGGTCCTCAGGGAACCAGGC 409126 5-10-5 447 1534 1553 1 ACCCTTGGTCACGCCGGCAT 410536 5-10-5 243 1527 1546 0 GCCACGCCGGCATCCCGGCC 410537 5-10-5 244 1528 1547 0 GGCCACGCCGGCATCCCGGC 410538 5-10-5 254 1539 1558 0 CTGGCACCCTTGGCCACGCC 410539 5-10-5 255 1540 1559 0 GCTGGCACCCTTGGCCACGC 410540 5-10-5 356 2005 2024 0 GCTGGTCCTCAGGGAACCAG 410562 5-10-5 411 2356 2375 0 ACTTTGCATTCCAGACCTGG 410563 5-10-5 413 2358 2377 0 TGACTTTGCATTCCAGACCT 410564 5-10-5 414 2359 2378 0 TTGACTTTGCATTCCAGACC 410565 5-10-5 419 2560 2579 0 CAGATGGCAACGGCTGTCAC 410566 5-10-5 420 2561 2580 0 GCAGATGGCAACGGCTGTCA 410567 5-10-5 421 2562 2581 0 AGCAGATGGCAACGGCTGTC 410568 5-10-5 422 2563 2582 0 CAGCAGATGGCAACGGCTGT 410569 5-10-5 423 2564 2583 0 GCAGCAGATGGCAACGGCTG 410570 5-10-5 424 2566 2585 0 CGGCAGCAGATGGCAACGGC 410571 5-10-5 425 2567 2586 0 CCGGCAGCAGATGGCAACGG 410572 5-10-5 427 2569 2588 0 CTCCGGCAGCAGATGGCAAC 410573 5-10-5 428 2570 2589 0 GCTCCGGCAGCAGATGGCAA 410733 5-10-5 231 1416 1435 0 AAGTCGGTGACCATGACCCT 410734 5-10-5 279 1749 1768 0 CCGGCAGCGGTGACCAGCAC 410737 5-10-5 313 1839 1858 0 GCAGGTGCTGCAGTCGCTGG 410738 5-10-5 324 1915 1934 0 GCTCGGCAGACAGCATCATG 410739 5-10-5 336 1963 1982 0 CAGAGAAGTGGATCAGTCTC 410740 5-10-5 345 1991 2010 0 AACCAGGCCTCATTGATGAC 410741 5-10-5 369 2116 2135 0 CCATCCGTGTAGGCCCCGAG 410753 5-10-5 215 1351 1370 0 ATACACCTCCACCAGGCTGC 410754 5-10-5 233 1428 1447 0 GGCACATTCTCGAAGTCGGT 410755 5-10-5 234 1463 1482 0 TGGCCTGTCTGTGGAAGCGG 410756 5-10-5 235 1490 1509 0 GGTGGGTGCCATGACTGTCA 410757 5-10-5 240 1515 1534 0 TCCCGGCCGCTGACCACCCC 410758 5-10-5 256 1545 1564 0 CGCATGCTGGCACCCTTGGC 410759 5-10-5 263 1594 1613 0 GGTGCCGCTAACCGTGCCCT 410760 5-10-5 265 1618 1637 0 CCGAATAAACTCCAGGCCTA 410761 5-10-5 271 1649 1668 0 GTGGCCCCACAGGCTGGACC 410762 5-10-5 272 1662 1681 0 AGCAGCACCACCAGTGGCCC 410763 5-10-5 273 1684 1703 0 GCTGTACCCACCCGCCAGGG 410764 5-10-5 274 1730 1749 0 CGACCCCAGCCCTCGCCAGG 410769 5-10-5 318 1858 1877 0 TCCCACTCTGTGACACAAAG 410770 5-10-5 327 1933 1952 0 CGGCCAGGGTGAGCTCCGGC 410771 5-10-5 360 2061 2080 0 ACCTGCCCCATGGGTGCTGG 410776 5-10-5 408 2245 2264 0 GTGCCAAGGTCCTCCACCTC 410777 5-10-5 409 2265 2284 0 TCAGCACAGGCGGCTTGTGG 410778 5-10-5 410 2295 2314 0 CCACGCACTGGTTGGGCTGA 410779 5-10-5 416 2375 2394 0 CGGGATTCCATGCTCCTTGA 410780 5-10-5 417 2405 2424 0 GCAGGCCACGGTCACCTGCT 410781 5-10-5 418 2442 2461 0 GGAGGGCACTGCAGCCAGTC 410782 5-10-5 429 2580 2599 0 CCAGGTGCCGGCTCCGGCAG 410783 5-10-5 430 2590 2609 0 GAGGCCTGCGCCAGGTGCCG

In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 3. In certain such embodiments, the nucleotide sequences illustrated in Table 12 have a 3-14-3 gap-widened motif. Table 14 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 3, having a 3-14-3 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 14 Gapmer antisense compounds having a 3-14-3 motif targeted to SEQ ID NO: 3 5′ Target 3′ Target Site on Site on SEQ ID SEQ ID SEQ ID Isis No Motif NO NO: 3 NO: 3 Mismatches Sequence (5′-3′) 399923 3-14-3 85 155 174 0 ACAAATTCCCAGACTCAGCA 399983 3-14-3 100 220 239 0 ATCTCAGGACAGGTGAGCAA 399984 3-14-3 116 234 253 0 GAGTAGAGATTCTCATCTCA 399924 3-14-3 129 290 309 0 GTGCCATCTGAACAGCACCT 399985 3-14-3 117 302 321 0 GAGTCTTCTGAAGTGCCATC 399986 3-14-3 81 377 396 0 AAGCAGGGCCTCAGGTGGAA 399925 3-14-3 110 400 419 0 CCTGGAACCCCTGCAGCCAG 399926 3-14-3 152 451 470 0 TTCAGGCAGGTTGCTGCTAG 399987 3-14-3 83 460 479 0 AAGGAAGACTTCAGGCAGGT 399927 3-14-3 140 472 491 0 TCAGCCAGGCCAAAGGAAGA 399988 3-14-3 137 586 605 0 TAGGGAGAGCTCACAGATGC 399928 3-14-3 136 596 615 0 TAGGAGAAAGTAGGGAGAGC 399929 3-14-3 132 653 672 0 TAAAAGCTGCAAGAGACTCA 399930 3-14-3 139 741 760 0 TCAGAGAAAACAGTCACCGA 399989 3-14-3 92 871 890 0 AGAGACAGGAAGCTGCAGCT 399931 3-14-3 142 878 897 0 TCATTTTAGAGACAGGAAGC 399932 3-14-3 113 915 934 0 GAATAACAGTGATGTCTGGC 399933 3-14-3 138 968 987 0 TCACAGCTCACCGAGTCTGC 399934 3-14-3 98 998 1017 0 AGTGTAAAATAAAGCCCCTA 399935 3-14-3 96 1075 1094 0 AGGACCCAAGTCATCCTGCT 399936 3-14-3 124 1105 1124 0 GGCCATCAGCTGGCAATGCT 399990 3-14-3 82 1144 1163 0 AAGGAAAGGGAGGCCTAGAG 399937 3-14-3 133 1149 1168 0 TAGACAAGGAAAGGGAGGCC 399938 3-14-3 103 1155 1174 0 ATTTCATAGACAAGGAAAGG 399939 3-14-3 155 1275 1294 0 CTTATAGTTAACACACAGAA 399991 3-14-3 156 1283 1302 0 AAGTCAACCTTATAGTTAAC 399940 3-14-3 146 1315 1334 0 TGACATTTGTGGGAGAGGAG 399941 3-14-3 161 1322 1341 0 TCCAAGGTGACATTTGTGGG 399882 3-14-3 19 1365 1384 0 CTGGTGTCTAGGAGATACAC 399953 3-14-3 20 1370 1389 0 GTATGCTGGTGTCTAGGAGA 399883 3-14-3 21 1390 1409 0 GATTTCCCGGTGGTCACTCT 399954 3-14-3 22 1396 1415 0 GCCCTCGATTTCCCGGTGGT 399955 3-14-3 23 1410 1429 0 GTGACCATGACCCTGCCCTC 399884 3-14-3 24 1420 1439 0 CTCGAAGTCGGTGACCATGA 399885 3-14-3 25 1453 1472 0 GTGGAAGCGGGTCCCGTCCT 399886 3-14-3 26 1500 1519 0 ACCCCTGCCAGGTGGGTGCC 399956 3-14-3 27 1505 1524 0 TGACCACCCCTGCCAGGTGG 399887 3-14-3 28 1534 1553 0 ACCCTTGGCCACGCCGGCAT 399888 3-14-3 29 1570 1589 0 TTGGCAGTTGAGCACGCGCA 399957 3-14-3 30 1575 1594 0 TTCCCTTGGCAGTTGAGCAC 399889 3-14-3 31 1607 1626 0 CCAGGCCTATGAGGGTGCCG 399890 3-14-3 32 1628 1647 0 GCTGGCTTTTCCGAATAAAC 399891 3-14-3 33 1740 1759 0 GTGACCAGCACGACCCCAGC 399897 3-14-3 39 1822 1841 0 TGGAGGCACCAATGATGTCC 399960 3-14-3 40 1833 1852 0 GCTGCAGTCGCTGGAGGCAC 399961 3-14-3 41 1844 1863 0 ACAAAGCAGGTGCTGCAGTC 399898 3-14-3 101 1898 1917 0 ATGGCTGCAATGCCAGCCAC 399962 3-14-3 42 1903 1922 0 GCATCATGGCTGCAATGCCA 399963 3-14-3 43 1911 1930 0 GGCAGACAGCATCATGGCTG 399964 3-14-3 44 1959 1978 0 GAAGTGGATCAGTCTCTGCC 399899 3-14-3 45 1967 1986 0 TTGGCAGAGAAGTGGATCAG 399965 3-14-3 46 1972 1991 0 CATCTTTGGCAGAGAAGTGG 399966 3-14-3 47 1978 1997 0 TGATGACATCTTTGGCAGAG 399967 3-14-3 48 1985 2004 0 GCCTCATTGATGACATCTTT 399968 3-14-3 49 1997 2016 0 TCAGGGAACCAGGCCTCATT 399900 3-14-3 50 2002 2021 0 GGTCCTCAGGGAACCAGGCC 399901 3-14-3 87 2009 2028 0 ACCCGCTGGTCCTCAGGGAA 399969 3-14-3 51 2016 2035 0 GGTCAGTACCCGCTGGTCCT 399902 3-14-3 119 2038 2057 0 GCAGGGCGGCCACCAGGTTG 399903 3-14-3 52 2073 2092 0 AAACAGCTGCCAACCTGCCC 399970 3-14-3 53 2078 2097 0 CTGCAAAACAGCTGCCAACC 399942 3-14-3 104 2085 2104 0 CACAGTCCTGCAAAACAGCT 399992 3-14-3 130 2095 2114 0 GTGCTGACCACACAGTCCTG 399904 3-14-3 54 2108 2127 0 GTAGGCCCCGAGTGTGCTGA 399971 3-14-3 55 2173 2192 0 AGAAACTGGAGCAGCTCAGC 399972 3-14-3 56 2179 2198 0 TCCTGGAGAAACTGGAGCAG 399908 3-14-3 60 2355 2374 0 CTTTGCATTCCAGACCTGGG 399973 3-14-3 61 2360 2379 0 CTTGACTTTGCATTCCAGAC 399909 3-14-3 62 2565 2584 0 GGCAGCAGATGGCAACGGCT 399910 3-14-3 154 2665 2684 0 TTTTAAAGCTCAGCCCCAGC 399974 3-14-3 63 2670 2689 0 AACCATTTTAAAGCTCAGCC 399911 3-14-3 64 2759 2778 0 TCAAGGGCCAGGCCAGCAGC 399975 3-14-3 65 2764 2783 0 CCCACTCAAGGGCCAGGCCA 399976 3-14-3 122 2837 2856 0 GGAGGGAGCTTCCTGGCACC 399912 3-14-3 66 2852 2871 0 ATGCCCCACAGTGAGGGAGG 399913 3-14-3 67 2861 2880 0 AATGGTGAAATGCCCCACAG 399914 3-14-3 153 2904 2923 0 TTGGGAGCAGCTGGCAGCAC 399915 3-14-3 68 3005 3024 0 CATGGGAAGAATCCTGCCTC 399916 3-14-3 69 3087 3106 0 GGAGATGAGGGCCATCAGCA 399917 3-14-3 70 3155 3174 0 TAGATGCCATCCAGAAAGCT 399977 3-14-3 71 3161 3180 0 TCTGGCTAGATGCCATCCAG 399918 3-14-3 72 3238 3257 0 GGCATAGAGCAGAGTAAAGG 399978 3-14-3 73 3243 3262 0 AGCCTGGCATAGAGCAGAGT 399979 3-14-3 135 3249 3268 0 TAGCACAGCCTGGCATAGAG 399919 3-14-3 112 3482 3501 0 GAAGAGGCTTGGCTTCAGAG 399980 3-14-3 74 3488 3507 0 AAGTAAGAAGAGGCTTGGCT 399920 3-14-3 75 3692 3711 0 GCTCAAGGAGGGACAGTTGT 399921 3-14-3 76 3727 3746 0 AAAGATAAATGTCTGCTTGC 399981 3-14-3 77 3732 3751 0 ACCCAAAAGATAAATGTCTG 399922 3-14-3 78 3798 3817 0 TCTTCAAGTTACAAAAGCAA 399982 3-14-3 99 3805 3824 0 ATAAATATCTTCAAGTTACA 405604 3-14-3 236 1493 1512 0 CCAGGTGGGTGCCATGACTG 405641 3-14-3 373 2170 2189 0 AACTGGAGCAGCTCAGCAGC 410583 3-14-3 243 1527 1546 0 GCCACGCCGGCATCCCGGCC 410584 3-14-3 244 1528 1547 0 GGCCACGCCGGCATCCCGGC 410585 3-14-3 245 1529 1548 0 TGGCCACGCCGGCATCCCGG 410586 3-14-3 246 1530 1549 0 TTGGCCACGCCGGCATCCCG 410587 3-14-3 247 1531 1550 0 CTTGGCCACGCCGGCATCCC 410588 3-14-3 248 1532 1551 0 CCTTGGCCACGCCGGCATCC 410589 3-14-3 249 1533 1552 0 CCCTTGGCCACGCCGGCATC 410590 3-14-3 250 1535 1554 0 CACCCTTGGCCACGCCGGCA 410591 3-14-3 251 1536 1555 0 GCACCCTTGGCCACGCCGGC 410592 3-14-3 252 1537 1556 0 GGCACCCTTGGCCACGCCGG 410593 3-14-3 253 1538 1557 0 TGGCACCCTTGGCCACGCCG 410594 3-14-3 254 1539 1558 0 CTGGCACCCTTGGCCACGCC 410595 3-14-3 255 1540 1559 0 GCTGGCACCCTTGGCCACGC 410596 3-14-3 348 1995 2014 0 AGGGAACCAGGCCTCATTGA 410597 3-14-3 349 1996 2015 0 CAGGGAACCAGGCCTCATTG 410598 3-14-3 350 1998 2017 0 CTCAGGGAACCAGGCCTCAT 410599 3-14-3 351 1999 2018 0 CCTCAGGGAACCAGGCCTCA 410600 3-14-3 352 2000 2019 0 TCCTCAGGGAACCAGGCCTC 410601 3-14-3 353 2001 2020 0 GTCCTCAGGGAACCAGGCCT 410602 3-14-3 354 2003 2022 0 TGGTCCTCAGGGAACCAGGC 410603 3-14-3 355 2004 2023 0 CTGGTCCTCAGGGAACCAGG 410604 3-14-3 356 2005 2024 0 GCTGGTCCTCAGGGAACCAG 410633 3-14-3 411 2356 2375 0 ACTTTGCATTCCAGACCTGG 410634 3-14-3 412 2357 2376 0 GACTTTGCATTCCAGACCTG 410635 3-14-3 413 2358 2377 0 TGACTTTGCATTCCAGACCT 410636 3-14-3 414 2359 2378 0 TTGACTTTGCATTCCAGACC 410637 3-14-3 419 2560 2579 0 CAGATGGCAACGGCTGTCAC 410638 3-14-3 420 2561 2580 0 GCAGATGGCAACGGCTGTCA 410639 3-14-3 421 2562 2581 0 AGCAGATGGCAACGGCTGTC 410640 3-14-3 422 2563 2582 0 CAGCAGATGGCAACGGCTGT 410641 3-14-3 423 2564 2583 0 GCAGCAGATGGCAACGGCTG 410642 3-14-3 424 2566 2585 0 CGGCAGCAGATGGCAACGGC 410643 3-14-3 425 2567 2586 0 CCGGCAGCAGATGGCAACGG 410644 3-14-3 426 2568 2587 0 TCCGGCAGCAGATGGCAACG 410645 3-14-3 427 2569 2588 0 CTCCGGCAGCAGATGGCAAC 410646 3-14-3 428 2570 2589 0 GCTCCGGCAGCAGATGGCAA

In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 3. In certain such embodiments, the nucleotide sequences illustrated in Table 12 have a 2-13-5 gap-widened motif. Table 15 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 3, having a 2-13-5 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 15 Gapmer antisense compounds having a 2-13-5 motif targeted to SEQ ID NO: 3 5′ Target 3′ Target Site to Site to SEQ ID SEQ ID SEQ ID ISIS No Motiff NO NO: 3 NO: 3 Mismatches Sequences (5′-3′) 410658 2-13-5 243 1527 1546 0 GCCACGCCGGCATCCCGGCC 410659 2-13-5 244 1528 1547 0 GGCCACGCCGGCATCCCGGC 410660 2-13-5 245 1529 1548 0 TGGCCACGCCGGCATCCCGG 410661 2-13-5 246 1530 1549 0 TTGGCCACGCCGGCATCCCG 410662 2-13-5 247 1531 1550 0 CTTGGCCACGCCGGCATCCC 410663 2-13-5 248 1532 1551 0 CCTTGGCCACGCCGGCATCC 410664 2-13-5 249 1533 1552 0 CCCTTGGCCACGCCGGCATC 410665 2-13-5 28 1534 1553 0 ACCCTTGGCCACGCCGGCAT 410666 2-13-5 250 1535 1554 0 CACCCTTGGCCACGCCGGCA 410667 2-13-5 251 1536 1555 0 GCACCCTTGGCCACGCCGGC 410668 2-13-5 252 1537 1556 0 GGCACCCTTGGCCACGCCGG 410669 2-13-5 253 1538 1557 0 TGGCACCCTTGGCCACGCCG 410670 2-13-5 254 1539 1558 0 CTGGCACCCTTGGCCACGCC 410671 2-13-5 255 1540 1559 0 GCTGGCACCCTTGGCCACGC 410672 2-13-5 348 1995 2014 0 AGGGAACCAGGCCTCATTGA 410673 2-13-5 349 1996 2015 0 CAGGGAACCAGGCCTCATTG 410674 2-13-5 49 1997 2016 0 TCAGGGAACCAGGCCTCATT 410675 2-13-5 350 1998 2017 0 CTCAGGGAACCAGGCCTCAT 410676 2-13-5 351 1999 2018 0 CCTCAGGGAACCAGGCCTCA 410677 2-13-5 352 2000 2019 0 TCCTCAGGGAACCAGGCCTC 410678 2-13-5 353 2001 2020 0 GTCCTCAGGGAACCAGGCCT 410679 2-13-5 50 2002 2021 0 GGTCCTCAGGGAACCAGGCC 410680 2-13-5 354 2003 2022 0 TGGTCCTCAGGGAACCAGGC 410681 2-13-5 355 2004 2023 0 CTGGTCCTCAGGGAACCAGG 410682 2-13-5 356 2005 2024 0 GCTGGTCCTCAGGGAACCAG 410713 2-13-5 60 2355 2374 0 CTTTGCATTCCAGACCTGGG 410714 2-13-5 411 2356 2375 0 ACTTTGCATTCCAGACCTGG 410715 2-13-5 412 2357 2376 0 GACTTTGCATTCCAGACCTG 410716 2-13-5 413 2358 2377 0 TGACTTTGCATTCCAGACCT 410717 2-13-5 414 2359 2378 0 TTGACTTTGCATTCCAGACC 410718 2-13-5 61 2360 2379 0 CTTGACTTTGCATTCCAGAC 410719 2-13-5 419 2560 2579 0 CAGATGGCAACGGCTGTCAC 410720 2-13-5 420 2561 2580 0 GCAGATGGCAACGGCTGTCA 410721 2-13-5 421 2562 2581 0 AGCAGATGGCAACGGCTGTC 410722 2-13-5 422 2563 2582 0 CAGCAGATGGCAACGGCTGT 410723 2-13-5 423 2564 2583 0 GCAGCAGATGGCAACGGCTG 410724 2-13-5 62 2565 2584 0 GGCAGCAGATGGCAACGGCT 410725 2-13-5 424 2566 2585 0 CGGCAGCAGATGGCAACGGC 410726 2-13-5 425 2567 2586 0 CCGGCAGCAGATGGCAACGG 410727 2-13-5 426 2568 2587 0 TCCGGCAGCAGATGGCAACG 410728 2-13-5 427 2569 2588 0 CTCCGGCAGCAGATGGCAAC 410729 2-13-5 428 2570 2589 0 GCTCCGGCAGCAGATGGCAA

The following embodiments set forth target regions of PCSK9 nucleic acids. Also illustrated are examples of antisense compounds targeted to the target regions. It is understood that the sequence set forth in each SEQ ID NO is independent of any modification to a sugar moiety, an internucleoside linkage, or a nucleobase. As such, antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an internucleoside linkage, or a nucleobase. Antisense compounds described by Isis Number (Isis No) indicate a combination of nucleobase sequence and motif.

In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 3: 220-253, 290-321, 377-419, 451-615, 653-672, 741-760, 871-897, 915-934, 968-1017, 1075-1124, 1075-1174, 1144-1174, 1275-1302, 1315-1341, 1351-1389, 1351-1447, 1365-1439, 1390-1417, 1390-1429, 1390-1439, 1399-1425, 1408-1435, 1420-1447, 1453-1482, 1490-1516, 1490-1564, 1500-1526, 1515-1541, 1527-1553, 1527-1554, 1528-1554, 1529-1555, 1529-1556, 1530-1556, 1530-1557, 1531-1557, 1532-1551, 1532-1558, 1533-1559, 1534-1559, 1535-1559, 1536-1559, 1537-1564, 1566-1602, 1566-1681, 1606-1626, 1618-1645, 1626-1653, 1684-1703, 1730-1781, 1740-1767, 1749-1775, 1758-1781, 1820-1847, 1820-1877, 1822-2198, 1830-1856, 1839-1865, 1840-1867, 1898-1924, 1898-2035, 1903-2127, 1907-1934, 1911-1938, 1946-1971, 1954-1980, 1959-2035, 1959-2057, 1963-1988, 1967-2035, 1972-1999, 1982-2008, 1991-2018, 1993-2019, 1995-2022, 1996-2023, 1997-2024, 1998-2025, 1999-2025, 2000-2025, 2009-2035, 2038-2139, 2061-2139, 2073-2099, 2078-2104, 2105-2131, 2112-2139, 2168-2198, 2170-2177, 2245-2284, 2295-2394, 2355-2381, 2355-2394, 2405-2461, 2560-2587, 2560-2609, 2561-2588, 2562-2589, 2563-2589, 2564-2589, 2565-2589, 2566-2589, 2567-2589, 2568-2589, 2665-2689, 2759-2783, 2837-2880, 2904-2923, 3005-3024, 3005-3174, 3083-3110, 3155-3184, 3238-3268, 3482-3711, or 3727-3751, or 3798-3824.

In certain embodiments, a target region is nucleotides 290-321 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 290-321 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 117 or 129. In certain such embodiments, an antisense compound targeted to nucleotides 290-321 of SEQ ID NO: 3 is selected from ISIS NOs: 395202, 399924, 399829 or 399985.

In certain embodiments, a target region is nucleotides 220-253 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 220-253 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 100 or 116. In certain such embodiments, an antisense compound targeted to nucleotides 220-253 of SEQ ID NO: 3 is selected from ISIS NOs: 399827, 399983, 399828 or 399984.

In certain embodiments, a target region is nucleotides 377-419 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 377-419 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 81 or 110. In certain such embodiments, an antisense compound targeted to nucleotides 377-419 of SEQ ID NO: 3 is selected from ISIS NOs: 399830, 399986, 395203 or 399925.

In certain embodiments, a target region is nucleotides 451-615 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 451-615 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 152, 140, 137 or 136. In certain such embodiments, an antisense compound targeted to nucleotides 451-615 of SEQ ID NO: 3 is selected from ISIS NOs: 395204, 399926, 399831, 399987, 395205, 399927, 399832, 399988, 395206 or 399928.

In certain embodiments, a target region is nucleotides 653-672 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 653-672 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 132. In certain such embodiments, an antisense compound targeted to nucleotides 653-672 of SEQ ID NO: 3 is selected from ISIS NOs: 395207 or 399929.

In certain embodiments, a target region is nucleotides 741-760 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 741-760 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 139. In certain such embodiments, an antisense compound targeted to nucleotides 741-760 of SEQ ID NO: 3 is selected from ISIS NOs: 395208 or 399930.

In certain embodiments, a target region is nucleotides 871-897 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 871-897 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 92 or 142. In certain such embodiments, an antisense compound targeted to nucleotides 871-897 of SEQ ID NO: 3 is selected from ISIS NOs: 399833, 399989, 395209 or 399931.

In certain embodiments, a target region is nucleotides 915-934 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 915-934 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 113. In certain such embodiments, an antisense compound targeted to nucleotides 915-934 of SEQ ID NO: 3 is selected from ISIS NOs: 395210 or 399932.

In certain embodiments, a target region is nucleotides 968-1017 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 968-1017 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 98 or 138. In certain such embodiments, an antisense compound targeted to nucleotides 968-1017 of SEQ ID NO: 3 is selected from ISIS NOs: 395211, 399933, 395212, or 399934.

In certain embodiments, a target region is nucleotides 1075-1124 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1075-1124 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 96 or 124. In certain such embodiments, an antisense compound targeted to nucleotides 1075-1124 of SEQ ID NO: 3 is selected from ISIS NOs: 395213, 399935, 395214, or 399936.

In certain embodiments, a target region is nucleotides 1075-1174 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1075-1174 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 82, 96, 103, 124, or 133. In certain such embodiments, an antisense compound targeted to nucleotides 1075-1174 of SEQ ID NO: 3 is selected from ISIS NOs: 395213, 399935, 395214, 399936, 399834, 399990, 395215, 399937, 395216, or 399938.

In certain embodiments, a target region is nucleotides 1144-1174 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1144-1174 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 82, 133 or 103. In certain such embodiments, an antisense compound targeted to nucleotides 1144-1174 of SEQ ID NO: 3 is selected from ISIS NOs: 399834, 399990, 395215, 399937, 395216 or 399938.

In certain embodiments, a target region is nucleotides 1275-1302 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1275-1302 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 155 or 156. In certain such embodiments, an antisense compound targeted to nucleotides 1275-1302 of SEQ ID NO: 3 is selected from ISIS NOs: 395217, 399939, 399835 or 399991.

In certain embodiments, a target region is nucleotides 1315-1341 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1315-1341 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 146 or 161. In certain such embodiments, an antisense compound targeted to nucleotides 1315-1341 of SEQ ID NO: 3 is selected from ISIS NOs: 395218, 399940, 395219 or 399941.

In certain embodiments, a target region is nucleotides 1351-1389 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1351-1389 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 215, 216, 19, 217 or 20. In certain such embodiments, an antisense compound targeted to nucleotides 1351-1389 of SEQ ID NO: 3 is selected from ISIS NOs: 410753, 406025, 395160, 399882, 406026, 399797 or 399953.

In certain embodiments, a target region is nucleotides 1351-1447 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1351-1447 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 215, 216, 19, 217, 20, 21, 218, 219, 220, 221, 22, 222, 223, 224, 225, 226, 227, 228, 23, 229, 230, 231, 232, 24, or 233. In certain such embodiments, an antisense compound targeted to nucleotides 1351-1447 of SEQ ID NO: 3 is selected from ISIS NOs: 410753, 406025, 395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, 399884 or 410754.

In certain embodiments, a target region is nucleotides 1365-1439 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1365-1439 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20, 21, 22, 23, 24, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, or 232. In certain such embodiments, an antisense compound targeted to nucleotides 1365-1439 of SEQ ID NO: 3 is selected from ISIS NOs: 395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, or 399884.

In certain embodiments, a target region is nucleotides 1390-1417 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1390-1417 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 218, 219, 220, 221, 22, 222 or 223. In certain such embodiments, an antisense compound targeted to nucleotides 1390-1417 of SEQ ID NO: 3 is selected from ISIS NOs: 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873 or 405874.

In certain embodiments, a target region is nucleotides 1390-1429 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1390-1429 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, or 228. In certain such embodiments, an antisense compound targeted to nucleotides 1390-1429 of SEQ ID NO: 3 is selected from ISIS NOs: 395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, or 399955.

In certain embodiments, a target region is nucleotides 1390-1439 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1390-1439 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 24, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, or 232. In certain such embodiments, an antisense compound targeted to nucleotides 1390-1439 of SEQ ID NO: 3 is selected from ISIS NOs: 395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, or 399884.

In certain embodiments, a target region is nucleotides 1399-1425 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1399-1425 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 224, 225, 226 or 227. In certain such embodiments, an antisense compound targeted to nucleotides 1399-1425 of SEQ ID NO: 3 is selected from ISIS NOs: 405875, 405876, 406027 or 406028.

In certain embodiments, a target region is nucleotides 1408-1435 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1408-1435 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 228, 23, 229, 230 or 231. In certain such embodiments, an antisense compound targeted to nucleotides 1408-1435 of SEQ ID NO: 3 is selected from ISIS NOs: 406029, 399799, 399955, 406030, 406031 or 410733.

In certain embodiments, a target region is nucleotides 1420-1447 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1420-1447 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 24 or 233. In certain such embodiments, an antisense compound targeted to nucleotides 1420-1447 of SEQ ID NO: 3 is selected from ISIS NOs: 395162, 399884 or 410754.

In certain embodiments, a target region is nucleotides 1453-1482 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1453-1482 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 25 or 234. In certain such embodiments, an antisense compound targeted to nucleotides 1453-1482 of SEQ ID NO: 3 is selected from ISIS NOs: 395163, 399885 or 410755.

In certain embodiments, a target region is nucleotides 1490-1516 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1490-1516 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 235, 236, or 237. In certain such embodiments, an antisense compound targeted to nucleotides 1490-1516 of SEQ ID NO: 3 is selected from ISIS NOs: 410756, 405604, or 406033.

In certain embodiments, a target region is nucleotides 1490-1564 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1490-1564 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 235, 236, 237, 26, 238, 27, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 447, 28, 250, 251, 252, 253, 254, 255, or 256. In certain such embodiments, an antisense compound targeted to nucleotides 1490-1564 of SEQ ID NO: 3 is selected from ISIS NOs: 410756, 405604, 406033, 395164, 399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, or 410758.

In certain embodiments, a target region is nucleotides 1500-1526 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1500-1526 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 238, 27, or 239. In certain such embodiments, an antisense compound targeted to nucleotides 1500-1526 of SEQ ID NO: 3 is selected from ISIS NOs: 395164, 399886, 406034, 399800, 399956, or 406035.

In certain embodiments, a target region is nucleotides 1515-1541 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1515-1541 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 240, 241, or 242. In certain such embodiments, an antisense compound targeted to nucleotides 1515-1541 of SEQ ID NO: 3 is selected from ISIS NOs: 399886, 406034, 399800, 399956, or 406035.

In certain embodiments, a target region is nucleotides 1527-1553 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1527-1553 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 243, 244, 245, 246, 247, 248, 447, 28 or 249. In certain such embodiments, an antisense compound targeted to nucleotides 1527-1553 of SEQ ID NO: 3 is selected from ISIS NOs: 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, or 410665.

In certain embodiments, a target region is nucleotides 1527-1554 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1527-1554 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 243, 244, 245, 246, 247, 248, 249, 447, 28, or 250. In certain such embodiments, an antisense compound targeted to nucleotides 1527-1554 of SEQ ID NO: 3 is selected from ISIS NOs: 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 405881, 410590, 410666, or 410665.

In certain embodiments, a target region is nucleotides 1528-1554 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1528-1554 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 244, 245, 246, 247, 248, 249, 447, 28, or 250. In certain such embodiments, an antisense compound targeted to nucleotides 1528-1554 of SEQ ID NO: 3 is selected from ISIS NOs: 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 409126, 410665, or 405881.

In certain embodiments, a target region is nucleotides 1529-1555 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1529-1555 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 245, 246, 247, 248, 249, 447, 28, 250 or 251. In certain such embodiments, an antisense compound targeted to nucleotides 1529-1555 of SEQ ID NO: 3 is selected from ISIS NOs: 406038, 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 409126, 410585, 410586, 410587, 410588, 410589, 410590, 410591, 410660, 410661, 410662, 410663, 410664, 410665, 410666, 410667, or 410667.

In certain embodiments, a target region is nucleotides 1529-1556 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1529-1556 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 245, 246, 247, 248, 249, 447, 28, 250, 251, or 252. In certain such embodiments, an antisense compound targeted to nucleotides 1529-1556 of SEQ ID NO: 3 is selected from ISIS NOs: 406038, 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 409126, 410585, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410660, 410661, 410662, 410663, 410664, 410665, 410666, 410667, or 410668.

In certain embodiments, a target region is nucleotides 1530-1556 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1530-1556 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 246, 247, 248, 249, 447, 28, 250, 251, or 252. In certain such embodiments, an antisense compound targeted to nucleotides 1530-1556 of SEQ ID NO: 3 is selected from ISIS NOs: 406038, 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 409126, 410585, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410660, 410661, 410662, 410663, 410664, 410665, 410666, 410667, 405884, 410593, 410669 or 410668.

In certain embodiments, a target region is nucleotides 1530-1557 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1530-1557 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 246, 247, 248, 249, 250, 251, 252, 253, or 447. In certain such embodiments, an antisense compound targeted to nucleotides 1530-1557 of SEQ ID NO: 3 is selected from ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 409126, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410593, 410661, 410662, 410663, 410664, 410665, 410666, 410667, 410668, or 410669.

In certain embodiments, a target region is nucleotides 1531-1557 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1531-1557 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 247, 248, 249, 250, 251, 252, 253, or 447. In certain such embodiments, an antisense compound targeted to nucleotides 1531-1557 of SEQ ID NO: 3 is selected from ISIS NOs: 395165, 399887, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 409126, 410587, 410588, 410589, 410590, 410591, 410592, 410593, 410662, 410663, 410664, 410665, 410666, 410667, 410668, or 410669.

In certain embodiments, a target region is nucleotides 1532-1551 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1532-1551 of SEQ ID NO: 3. In one such embodiment, an antisense compound targeted to nucleotides 1532-1551 of SEQ ID NO: 3 is ISIS NO: 405879.

In certain embodiments, a target region is nucleotides 1532-1558 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1532-1558 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 248, 249, 250, 251, 252, 253, 254 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 1532-1558 of SEQ ID NO: 3 is selected from ISIS NOs: 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, or 410670.

In certain embodiments, a target region is nucleotides 1533-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1533-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 249, 250, 251, 252, 253, 254, 255 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 1533-1559 of SEQ ID NO: 3 is selected from ISIS NOs: 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.

In certain embodiments, a target region is nucleotides 1534-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1534-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 250, 251, 252, 253, 254, 255 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 1534-1559 of SEQ ID NO: 3 is selected from ISIS NOs: 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, or 410671.

In certain embodiments, a target region is nucleotides 1535-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1535-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 250, 251, 252, 253, 254, 255 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 1535-1559 of SEQ ID NO: 3 is selected from ISIS NOs: 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594 or 410671.

In certain embodiments, a target region is nucleotides 1536-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1536-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 251, 252, 253, 254, or 255. In certain such embodiments, an antisense compound targeted to nucleotides 1536-1559 of SEQ ID NO: 3 is selected from ISIS NOs: 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.

In certain embodiments, a target region is nucleotides 1537-1564 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1537-1564 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 251, 252, 253, 254, 255, or 256. In certain such embodiments, an antisense compound targeted to nucleotides 1537-1564 of SEQ ID NO: 3 is selected from ISIS NOs: 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, 410671, or 410758.

In certain embodiments, a target region is nucleotides 1566-1602 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1566-1602 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 29, 30, 31, 32, 257, 258, 259, 260, 261, or 262. In certain such embodiments, an antisense compound targeted to nucleotides 1566-1602 of SEQ ID NO: 3 is selected from ISIS NOs: 395166, 395167, 395168, 399801, 399888, 399889, 399890, 399957, 405909, 405910, 405911, 405912, 406039, 406040, 406041, 406042, 406043, or 406044.

In certain embodiments, a target region is nucleotides 1566-1681 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1566-1681 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 29, 30, 31, 32, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, or 272. In certain such embodiments, an antisense compound targeted to nucleotides 1566-1681 of SEQ ID NO: 3 is selected from ISIS NOs: 395166, 395167, 395168, 399801, 399888, 399889, 399890, 399957, 405909, 405910, 405911, 405912, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 408642, 410759, 410760, 410761, or 410762.

In certain embodiments, a target region is nucleotides 1606-1626 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1606-1626 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 264 or 31. In certain such embodiments, an antisense compound targeted to nucleotides 1606-1626 of SEQ ID NO: 3 is selected from ISIS NOs: 408642, 395167, or 399889.

In certain embodiments, a target region is nucleotides 1618-1645 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1618-1645 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 265 or 266. In certain such embodiments, an antisense compound targeted to nucleotides 1618-1645 of SEQ ID NO: 3 is selected from ISIS NOs: 410760 or 406045.

In certain embodiments, a target region is nucleotides 1626-1653 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1626-1653 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 32, 266, 267, 268, or 269. In certain such embodiments, an antisense compound targeted to nucleotides 1626-1653 of SEQ ID NO: 3 is selected from ISIS NOs: 395168, 399890, 405909, 405910, 405911, or 406045.

In certain embodiments, a target region is nucleotides 1684-1703 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1684-1703 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 273. In certain such embodiments, an antisense compound targeted to nucleotides 1684-1703 of SEQ ID NO: 3 is selected from ISIS NOs: 410763.

In certain embodiments, a target region is nucleotides 1730-1781 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1730-1781 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 33, 33, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, or 285. In certain such embodiments, an antisense compound targeted to nucleotides 1730-1781 of SEQ ID NO: 3 is selected from ISIS NOs: 395169, 399891, 405913, 405914, 405915, 405916, 405917, 405918, 405919, 405920, 405921, 405922, 410734, or 410764.

In certain embodiments, a target region is nucleotides 1740-1767 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1740-1767 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 33, 275, 276, 277, or 278. In certain such embodiments, an antisense compound targeted to nucleotides 1740-1767 of SEQ ID NO: 3 is selected from ISIS NOs: 395169, 399891, 405913, 405914, 405915, or 405916.

In certain embodiments, a target region is nucleotides 1749-1775 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1749-1775 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 279, 280, 281 or 282. In certain such embodiments, an antisense compound targeted to nucleotides 1749-1775 of SEQ ID NO: 3 is selected from ISIS NOs: 410734, 405917, 405918 or 405919.

In certain embodiments, a target region is nucleotides 1758-1781 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1758-1781 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 283, 284, or 285. In certain such embodiments, an antisense compound targeted to nucleotides 1758-1781 of SEQ ID NO: 3 is selected from ISIS NOs: 405920, 405921, or 405922.

In certain embodiments, a target region is nucleotides 1820-1847 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1820-1847 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 39, 306, 307, 308, or 309. In certain such embodiments, an antisense compound targeted to nucleotides 1820-1847 of SEQ ID NO: 3 is selected from ISIS NOs: 395175, 399897, 405937, 405938, 405939, or 405940.

In certain embodiments, a target region is nucleotides 1820-1877 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1820-1877 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 39, 40, 41, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, or 318. In certain such embodiments, an antisense compound targeted to nucleotides 1820-1877 of SEQ ID NO: 3 is selected from ISIS NOs: 395175, 399804, 399805, 399897, 399960, 399961, 405937, 405938, 405939, 405940, 405941, 405942, 405943, 405944, 405945, 405946, 405947, 410737, or 410769.

In certain embodiments, a target region is nucleotides 1822-2198 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1822-2198 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 39, 307, 308, 309, 310, 40, 311, 312, 313, 314, 315, 41, 41, 316, 317, 318, 101, 319, 42, 320, 321, 322, 43, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 44, 335, 336, 337, 45, 338, 46, 339, 340, 47, 341, 342, 48, 343, 344, 345, 346, 347, 3, or 366. In certain such embodiments, an antisense compound targeted to nucleotides 1822-2198 of SEQ ID NO: 3 is selected from ISIS NOs: 395175, 399897, 405938, 405939, 405940, 405941, 399804, 399960, 405942, 405943, 410737, 405944, 405945, 399805, 399961, 405946, 405947, 410769, 395176, 399898, 405948, 399806, 399962, 405949, 405950, 405951, 399807, 399963, 405952, 410738, 405953, 405954, or 405979.

In certain embodiments, a target region is nucleotides 1830-1856 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1830-1856 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 40, 310, 311, or 312. In certain such embodiments, an antisense compound targeted to nucleotides 1830-1856 of SEQ ID NO: 3 is selected from ISIS NOs: 399804, 399960, 405941, 405942, or 405943.

In certain embodiments, a target region is nucleotides 1839-1865 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1839-1865 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 41, 313, 314, 315, or 316. In certain such embodiments, an antisense compound targeted to nucleotides 1839-1865 of SEQ ID NO: 3 is selected from ISIS NOs: 399805, 399961, 405944, 405945, 405946, or 410737.

In certain embodiments, a target region is nucleotides 1840-1867 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1840-1867 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 314, 315, 316, or 317. In certain such embodiments, an antisense compound targeted to nucleotides 1840-1867 of SEQ ID NO: 3 is selected from ISIS NOs: 399961, 405944, 405945, 405946, 410737, or 405947.

In certain embodiments, a target region is nucleotides 1898-1924 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1898-1924 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 42, 101, 319, or 320. In certain such embodiments, an antisense compound targeted to nucleotides 1898-1924 of SEQ ID NO: 3 is selected from ISIS NOs: 395176, 399898, 405948, 399806, 399962, 405949, or 405949.

In certain embodiments, a target region is nucleotides 1898-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1898-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 87, 101, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, or 359. In certain such embodiments, an antisense compound targeted to nucleotides 1898-2035 of SEQ ID NO: 3 is selected from ISIS NOs: 395176, 395177, 395178, 395179, 399806, 399807, 399808, 399809, 399810, 399811, 399812, 399813, 399898, 399899, 399900, 399901, 399962, 399963, 399964, 399965, 399966, 399967, 399968, 399969, 405885, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 405948, 405949, 405950, 405951, 405952, 405953, 405954, 405955, 405956, 405957, 405958, 405959, 405960, 405961, 405962, 405963, 405964, 405965, 405966, 405967, 405968, 405969, 405970, 405971, 405972, 405973, 405974, 408653, 410540, 410596, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681, 410682, 410738, 410739, 410740, or 410770.

In certain embodiments, a target region is nucleotides 1903-2127 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1903-2127 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 42, 320, 321, 322, 43, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 44, 335, 336, 337, 45, 338, 46, 339, 340, 47, 341, 342, 48, 343, 344, 345, 346, 347, 348, 349, 49, 350, 351, 352, 353, 50, 354, 355, 356, 357, 87, 358, 359, 51, 119, 360, or 54. In certain such embodiments, an antisense compound targeted to nucleotides 1903-2127 of SEQ ID NO: 3 is selected from ISIS NOs: 399806, 399962, 405949, 405950, 405951, 399807, 399963, 405952, 410738, 405953, 405954, 410770, 405955, 405956, 405957, 405958, 405959, 405960, 405961, 399808, 399964, 405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, 405965, 405966, 399810, or 399967.

In certain embodiments, a target region is nucleotides 1907-1934 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1907-1934 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 321, 322, 323, or 324. In certain such embodiments, an antisense compound targeted to nucleotides 1907-1934 of SEQ ID NO: 3 is selected from ISIS NOs: 405950, 405951, 399807, 399963, 405952, or 410738.

In certain embodiments, a target region is nucleotides 1911-1938 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1911-1938 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 323, 324, 325, or 326. In certain such embodiments, an antisense compound targeted to nucleotides 1911-1938 of SEQ ID NO: 3 is selected from ISIS NOs: 399807, 399963, 405952, 410738, 405953, or 405954.

In certain embodiments, a target region is nucleotides 1946-1971 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1946-1971 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 328, 329, 330, or 331. In certain such embodiments, an antisense compound targeted to nucleotides 1946-1971 of SEQ ID NO: 3 is selected from ISIS NOs: 405955, 405956, 405957, or 405958.

In certain embodiments, a target region is nucleotides 1954-1980 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1954-1980 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 332, 333, 334, or 335. In certain such embodiments, an antisense compound targeted to nucleotides 1954-1980 of SEQ ID NO: 3 is selected from ISIS NOs: 405959, 405960, 405961, 399808, 399964, or 405962.

In certain embodiments, a target region is nucleotides 1959-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1959-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 335, 336, 337, 45, 338, 46, 339, 340, 47, 341, 342, 48, 343, 344, 345, 346, 347, 348, 349, 49, 350, 351, 352, 353, 50, 354, 355, 356, 357, 87, 358, 359, or 51. In certain such embodiments, an antisense compound targeted to nucleotides 1959-2035 of SEQ ID NO: 3 is selected from ISIS NOs: 399808, 399964, 405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, 405965, 405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969, 405970, 410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, or 399969.

In certain embodiments, a target region is nucleotides 1959-2057 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1959-2057 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 335, 336, 337, 45, 338, 46, 339, 340, 47, 341, 342, 48, 343, 344, 345, 346, 347, 348, 349, 49, 350, 351, 352, 353, 50, 354, 355, 356, 357, 87, 358, 359, 51, or 119. In certain such embodiments, an antisense compound targeted to nucleotides 1959-2057 of SEQ ID NO: 3 is selected from ISIS NOs: 399808, 399964, 405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, 405965, 405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969, 405970, 410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, or 399902.

In certain embodiments, a target region is nucleotides 1963-1988 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1963-1988 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 45, 336, 337 or 338. In certain such embodiments, an antisense compound targeted to nucleotides 1963-1988 of SEQ ID NO: 3 is selected from ISIS NOs: 410739, 405963, 395177, 399899, or 405964.

In certain embodiments, a target region is nucleotides 1967-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1967-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 45, 338, 46, 339, 340, 47, 341, 342, 48, 343, 344, 345, 346, 347, 348, 349, 49, 350, 351, 352, 353, 50, 354, 355, 356, 357, 87, 358, 359 or 51. In certain such embodiments, an antisense compound targeted to nucleotides 1967-2035 of SEQ ID NO: 3 is selected from ISIS NOs: 395177, 399899, 405964, 399809, 399965, 405965, 405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969, 405970, 410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, or 399969.

In certain embodiments, a target region is nucleotides 1972-1999 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1972-1999 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 46, 47, 339, 340, or 341. In certain such embodiments, an antisense compound targeted to nucleotides 1972-1999 of SEQ ID NO: 3 is selected from ISIS NOs: 399809, 399965, 405965, 405966, 399810, 399966, or 405967.

In certain embodiments, a target region is nucleotides 1982-2008 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1982-2008 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 48, 342, 343, or 344. In certain such embodiments, an antisense compound targeted to nucleotides 1982-2008 of SEQ ID NO: 3 is selected from ISIS NOs: 405968, 399811, 399967, 405969, or 405970.

In certain embodiments, a target region is nucleotides 1991-2018 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1991-2018 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 345, 346, 347, 348, 349, 350, or 351. In certain such embodiments, an antisense compound targeted to nucleotides 1991-2018 of SEQ ID NO: 3 is selected from ISIS NOs: 399812, 399968, 405885, 405886, 405887, 405888, 405889, 405890, 410596, 410597, 410598, 410599, 410672, 410673, 410674, 410675, 410676, or 410740.

In certain embodiments, a target region is nucleotides 1993-2019 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1993-2019 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 346, 347, 348, 349, 350, or 354. In certain such embodiments, an antisense compound targeted to nucleotides 1993-2019 of SEQ ID NO: 3 is selected from ISIS NOs: 405885, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, or 410677.

In certain embodiments, a target region is nucleotides 1995-2022 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1995-2022 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 348, 349, 350, 351, 352, 353, 50, or 351 In certain such embodiments, an antisense compound targeted to nucleotides 1995-2022 of SEQ ID NO: 3 is selected from ISIS NOs: 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 405892, 410601, 410678, 395178, 399900, 410679, 408653, 410602, or 410680.

In certain embodiments, a target region is nucleotides 1996-2023 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1996-2023 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 349, 350, 351, 352, 353, 354, or 355. In certain such embodiments, an antisense compound targeted to nucleotides 1996-2023 of SEQ ID NO: 3 is selected from ISIS NOs: 395178, 399812, 399900, 399968, 405888, 405889, 405890, 405891, 405892, 405971, 408653, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680, or 410681.

In certain embodiments, a target region is nucleotides 1997-2024 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1997-2024 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 350, 351, 352, 353, 354, 355, or 356. In certain such embodiments, an antisense compound targeted to nucleotides 1997-2024 of SEQ ID NO: 3 is selected from ISIS NOs: 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, or 410682.

In certain embodiments, a target region is nucleotides 1998-2025 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1998-2025 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 50, 350, 351, 352, 353, 354, 355, 356 or 357. In certain such embodiments, an antisense compound targeted to nucleotides 1998-2025 of SEQ ID NO: 3 is selected from ISIS NOs: 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.

In certain embodiments, a target region is nucleotides 1999-2025 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1999-2025 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 50, 351, 352, 353, 354, 355, 356, or 357. In certain such embodiments, an antisense compound targeted to nucleotides 1999-2025 of SEQ ID NO: 3 is selected from ISIS NOs: 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.

In certain embodiments, a target region is nucleotides 2000-2025 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2000-2025 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 351, 352, 353, 354, 355, 356, or 357. In certain such embodiments, an antisense compound targeted to nucleotides 2000-2025 of SEQ ID NO: 3 is selected from ISIS NOs: 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.

In certain embodiments, a target region is nucleotides 2009-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2009-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 51, 87, 358, or 359. In certain such embodiments, an antisense compound targeted to nucleotides 2009-2035 of SEQ ID NO: 3 is selected from ISIS NOs: 395179, 399813, 399901, 399969, 405973, or 405974.

In certain embodiments, a target region is nucleotides 2038-2139 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2038-2139 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 52, 53, 54, 104, 119, 130, 360, 361, 362, 363, 365, 366, 367, 368, 369, 370, or 371. In certain such embodiments, an antisense compound targeted to nucleotides 2038-2139 of SEQ ID NO: 3 is selected from ISIS NOs: 395180, 395181, 395182, 395220, 399814, 399836, 399902, 399903, 399904, 399942, 399970, 399992, 405975, 405976, 405977, 405978, 405979, 405980, 405981, 405982, 405983, 410741, or 410771.

In certain embodiments, a target region is nucleotides 2061-2139 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2061-2139 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 52, 53, 54, 104, 130, 360, 361, 362, 363, 365, 366, 367, 368, 369, 370, or 371. In certain such embodiments, an antisense compound targeted to nucleotides 2061-2139 of SEQ ID NO: 3 is selected from ISIS NOs: 395181, 395182, 395220, 399814, 399836, 399903, 399904, 399942, 399970, 399992, 405975, 405976, 405977, 405978, 405979, 405980, 405981, 405982, 405983, 410741, or 410771.

In certain embodiments, a target region is nucleotides 2073-2099 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2073-2099 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 52, 53, 361, or 362. In certain such embodiments, an antisense compound targeted to nucleotides 2073-2099 of SEQ ID NO: 3 is selected from ISIS NOs: 395181, 399814, 399903, 399970, 405975, or 405976.

In certain embodiments, a target region is nucleotides 2078-2104 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2078-2104 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 53, 104, 362, or 363. In certain such embodiments, an antisense compound targeted to nucleotides 2078-2104 of SEQ ID NO: 3 is selected from ISIS NOs: 395220, 399814, 399942, 399970, 405976, or 405977.

In certain embodiments, a target region is nucleotides 2105-2131 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2105-2131 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 54, 365, 366, or 367. In certain such embodiments, an antisense compound targeted to nucleotides 2105-2131 of SEQ ID NO: 3 is selected from ISIS NOs: 395182, 399904, 405978, 405979, or 405980.

In certain embodiments, a target region is nucleotides 2112-2139 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2112-2139 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 367, 368, 369, 370, or 371. In certain such embodiments, an antisense compound targeted to nucleotides 2112-2139 of SEQ ID NO: 3 is selected from ISIS NOs: 405980, 405981, 410741, 405982, or 405983.

In certain embodiments, a target region is nucleotides 2168-2198 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2168-2198 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 56, 372, 373, 374, or 375. In certain such embodiments, an antisense compound targeted to nucleotides 2168-2198 of SEQ ID NO: 3 is selected from ISIS NOs: 399815, 399816, 399971, 399972, 405641, 405984, 405985, 405986, or 405987.

In certain embodiments, a target region is nucleotides 2170-2177 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2170-2177 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 373, 374, or 375. In certain such embodiments, an antisense compound targeted to nucleotides 2170-2177 of SEQ ID NO: 3 is selected from ISIS NOs: 399815, 399971, 405641, 405985, 405986, or 405987.

In certain embodiments, a target region is nucleotides 2245-2284 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2245-2284 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 408 or 409. In certain such embodiments, an antisense compound targeted to nucleotides 2245-2284 of SEQ ID NO: 3 is selected from ISIS NOs: 410776 or 410777.

In certain embodiments, a target region is nucleotides 2295-2394 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2295-2394 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 410, 411, 412, 413, 414, 415, or 416. In certain such embodiments, an antisense compound targeted to nucleotides 2295-2394 of SEQ ID NO: 3 is selected from ISIS NOs: 395186, 399817, 399908, 399973, 405996, 405997, 410562, 410563, 410564, 410633, 410634, 410635, 410636, 410713, 410714, 410715, 410716, 410717, 410718, 410778, or 410779.

In certain embodiments, a target region is nucleotides 2355-2381 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2355-2381 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413, 414, or 415. In certain such embodiments, an antisense compound targeted to nucleotides 2355-2381 of SEQ ID NO: 3 is selected from ISIS NOs: 395186, 399817, 399908, 399973, 405996, 405997, 410562, 410563, 410564, 410633, 410634, 410635, 410636, 410713, 410714, 410715, 410716, 410717, or 410718.

In certain embodiments, a target region is nucleotides 2355-2394 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2355-2394 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413, 414, 415, or 416. In certain such embodiments, an antisense compound targeted to nucleotides 2355-2394 of SEQ ID NO: 3 is selected from ISIS NOs: 395186, 399817, 399908, 399973, 405996, 405997, 410562, 410563, 410564, 410633, 410634, 410635, 410636, 410713, 410714, 410715, 410716, 410717, 410718, or 410779.

In certain embodiments, a target region is nucleotides 2405-2461 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2405-2461 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 417 or 418. In certain such embodiments, an antisense compound targeted to nucleotides 2405-2461 of SEQ ID NO: 3 is selected from ISIS NOs: 410780 or 410781.

In certain embodiments, a target region is nucleotides 2560-2587 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2560-2587 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422, 423, 424, 425, or 426. In certain such embodiments, an antisense compound targeted to nucleotides 2560-2587 of SEQ ID NO: 3 is selected from ISIS NOs: 395187, 399909, 405998, 410565, 410566, 410567, 410568, 410569, 410570, 410571, 410637, 410638, 410639, 410640, 410641, 410642, 410643, 410644, 410719, 410720, 410721, 410722, 410723, 410724, 410725, 410726, or 410727.

In certain embodiments, a target region is nucleotides 2560-2609 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2560-2609 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, or 430. In certain such embodiments, an antisense compound targeted to nucleotides 2560-2609 of SEQ ID NO: 3 is selected from ISIS NOs: 395187, 399909, 405998, 410565, 410566, 410567, 410568, 410569, 410570, 410571, 410572, 410573, 410637, 410638, 410639, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410719, 410720, 410721, 410722, 410723, 410724, 410725, 410726, 410727, 410728, or 410783.

In certain embodiments, a target region is nucleotides 2561-2588 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2561-2588 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 420, 421, 422, 423, 424, 425, 426, 427, or 62. In certain such embodiments, an antisense compound targeted to nucleotides 2561-2588 of SEQ ID NO: 3 is selected from ISIS NOs: 410566, 410638, 410720, 410567, 410639, 410721, 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572, 410645, or 410728.

In certain embodiments, a target region is nucleotides 2562-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2562-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 421, 422, 423, 424, 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2562-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 395187, 399909, 405998, 410567, 410568, 410569, 410570, 410571, 410572, 410573, 410639, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410721, 410722, 410723, 410724, 410725, 410726, 410727, 410728, or 410729.

In certain embodiments, a target region is nucleotides 2563-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2563-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 422, 423, 424, 425, 426, 427, 428 or 62. In certain such embodiments, an antisense compound targeted to nucleotides 2563-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2564-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2564-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 423, 424, 425, 426, 427, 428 or 62. In certain such embodiments, an antisense compound targeted to nucleotides 2564-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2565-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2565-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 424, 425, 426, 427, 428 or 62. In certain such embodiments, an antisense compound targeted to nucleotides 2565-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2566-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2566-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 424, 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2566-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 410570, 410642, 410725, 410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2567-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2567-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2567-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2568-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2568-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 426, 427 or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2568-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 405988, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.

In certain embodiments, a target region is nucleotides 2665-2689 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2665-2689 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 63 or 154. In certain such embodiments, an antisense compound targeted to nucleotides 2665-2689 of SEQ ID NO: 3 is selected from ISIS NOs: 395188, 399910, 399818, or 399974.

In certain embodiments, a target region is nucleotides 2759-2783 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2759-2783 SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 64 or 65. In certain such embodiments, an antisense compound targeted to nucleotides 2759-2783 of SEQ ID NO: 3 is selected from ISIS NOs: 395189, 399911, 399819, or 399975.

In certain embodiments, a target region is nucleotides 2837-2880 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2837-2880 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 66, 67, or 122. In certain such embodiments, an antisense compound targeted to nucleotides 2837-2880 of SEQ ID NO: 3 is selected from ISIS NOs: 399820, 399976, 395190, 399912, 395191, or 399913.

In certain embodiments, a target region is nucleotides 2904-2923 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2904-2923 SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 153. In certain such embodiments, an antisense compound targeted to nucleotides 2904-2923 of SEQ ID NO: 3 is selected from ISIS NOs: 395192 or 399914.

In certain embodiments, a target region is nucleotides 3005-3024 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3005-3024 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 68. In certain such embodiments, an antisense compound targeted to nucleotides 3005-3024 of SEQ ID NO: 3 is selected from ISIS NOs: 395193 or 399915.

In certain embodiments, a target region is nucleotides 3005-3174 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3005-3174 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 68, 431, 432, 433, 434, 69, 435, 436, 437, 438 or 70. In certain such embodiments, an antisense compound targeted to nucleotides 3005-3174 of SEQ ID NO: 3 is selected from ISIS NOs: 395193, 399915, 405893, 405894, 405895, 405896, 395194, 399916, 405897, 405898, 405899, 405900, 395195, or 399917.

In certain embodiments, a target region is nucleotides 3083-3110 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3083-3110 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 69, 69, 431, 432, 433, 434, 435, 436, 437, or 438. In certain such embodiments, an antisense compound targeted to nucleotides 3083-3110 of SEQ ID NO: 3 is selected from ISIS NOs: 395194, 399916, 405893, 405894, 405895, 405896, 405897, 405898, 405899, or 405900.

In certain embodiments, a target region is nucleotides 3155-3184 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3155-3184 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441, 442, 443, 444, 445, or 446. In certain such embodiments, an antisense compound targeted to nucleotides 3155-3184 of SEQ ID NO: 3 is selected from ISIS NOs: 395195, 399821, 399917, 399977, 405901, 405902, 405903, 405904, 405905, 405906, 405907, or 405908.

In certain embodiments, a target region is nucleotides 3238-3268 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3238-3268 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 72, 73, or 135. In certain such embodiments, an antisense compound targeted to nucleotides 3238-3268 of SEQ ID NO: 3 is selected from ISIS NOs: 395196, 399822, 399823, 399918, 399978, or 399979.

In certain embodiments, a target region is nucleotides 3482-3711 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3482-3711 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 74, 75, or 112. In certain such embodiments, an antisense compound targeted to nucleotides 3482-3711 of SEQ ID NO: 3 is selected from ISIS NOs: 395197, 395198, 399824, 399919, 399920, or 399980.

In certain embodiments, a target region is nucleotides 3727-3751 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3727-3751 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 76 or 77. In certain such embodiments, an antisense compound targeted to nucleotides 3727-3751 of SEQ ID NO: 3 is selected from ISIS NOs: 395199, 399921, 399825, or 399981.

In certain embodiments, a target region is nucleotides 3798-3824 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3798-3824 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 78 or 99. In certain such embodiments, an antisense compound targeted to nucleotides 3798-3824 of SEQ ID NO: 3 is selected from ISIS NOs: 395200, 399922, 399826, or 399982.

In certain embodiments, short gapmer antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gapmer antisense compounds are targeted to SEQ ID NO: 1, SEQ ID NO: 2 and/or SEQ ID NO: 3. In certain such embodiments, the nucleotide sequences illustrated in Table 15.1 have a 2-10-2 gapmer motif. Table 15.1 illustrates gapmer antisense compounds targeted to SEQ ID NO: 1, SEQ ID NO: 2 and/or SEQ ID NO: 3, having a 2-10-2 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.

TABLE 15.1 Gapmer antisense compounds having a 2-10-2 motif targeted to SEQ ID NO: 1, SEQ ID NO: 2 and/or SEQ ID NO: 3 5′ 3′ Target Target Site Site Target on on SEQ SEQ ID ISIS SEQ ID SEQ ID ID NO NO Motif OligoSeq NO: 1 NO: 1 NO 1 406539 2-10-2 GCCTATGAGGGTGC 1079 1092 458 1 406540 2-10-2 TCCAGGCCTATGAG 1084 1097 459 1 406551 2-10-2 CAGCTCAGCAGCTC 1735 1748 460 1 406595 2-10-2 TTAATCAGGGAGCC 2877 2890 461 2 406551 2-10-2 CAGCTCAGCAGCTC 21181 21194 460 2 406595 2-10-2 TTAATCAGGGAGCC 26684 26697 461 3 406539 2-10-2 GCCTATGAGGGTGC 1609 1622 458 3 406540 2-10-2 TCCAGGCCTATGAG 1614 1627 459 3 406551 2-10-2 CAGCTCAGCAGCTC 2168 2181 460 3 406595 2-10-2 TTAATCAGGGAGCC 3132 3145 461

In a certain embodiment, a short antisense compound has increased potency compared to a 20 nucleotide compound. In a certain embodiment such short antisense compound and 20 nucleotide compound target the same region. In a certain embodiment, a short antisense compound has increased potency compared to a 20 nucleotide compound containing the sequence of the short antisense compound.

In certain embodiments, the desired effect is a reduction in mRNA target nucleic acid levels. In other embodiments, the desired effect is reduction of levels of protein encoded by the target nucleic acid or a phenotypic change associated with the target nucleic acid.

In one embodiment, a target region is a structurally defined region of the nucleic acid. For example, a target region may encompass a 3′ UTR, a 5′ UTR, an exon, an intron, a coding region, a translation initiation region, translation termination region, or other defined nucleic acid region. In other embodiments, a target region may encompass the sequence from a 5′ target site of one target segment within the target region to a 3′ target site of another target segment within the target region.

Targeting includes determination of at least one target segment to which an antisense compound hybridizes, such that a desired effect occurs. A target region may contain one or more target segments. Multiple target segments within a target region may be overlapping. Alternatively, they may be non-overlapping. In one embodiment, target segments within a target region are separated by no more than about 10 nucleotides on the target nucleic acid. In another embodiment, target segments within a target region are separated by no more than about 5 nucleotides on the target nucleic acid. In additional embodiments, target segments are contiguous.

Suitable target segments may be found within a 5′ UTR, a coding region, a 3′ UTR, an intron, or an exon. Target segments containing a start codon or a stop codon are also suitable target segments.

The determination of suitable target segments may include a comparison of the sequence of a target nucleic acid to other sequences throughout the genome. For example, the BLAST algorithm may be used to identify regions of similarity amongst different nucleic acids. This comparison can prevent the selection of antisense compound sequences that may hybridize in a non-specific manner to sequences other than a selected target nucleic acid (i.e., non-target or off-target sequences).

There may be variation in activity (e.g., as defined by percent reduction of target nucleic acid levels) of the antisense compounds within an active target region. In one embodiment, reductions in PCSK9 mRNA levels are indicative of inhibition of PCSK9 expression. Reductions in levels of a PCSK9 protein are also indicative of inhibition of target mRNA expression. Further, phenotypic changes are indicative of inhibition of PCSK9 expression. For example, a decrease in LDL-C levels is indicative of inhibition of PCSK9 expression.

Hybridization

For example, hybridization may occur between an antisense compound disclosed herein and a PCSK9 nucleic acid. The most common mechanism of hybridization involves hydrogen bonding (e.g., Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding) between complementary nucleobases of the nucleic acid molecules.

Hybridization can occur under varying conditions. Stringent conditions are sequence-dependent and are determined by the nature and composition of the nucleic acid molecules to be hybridized.

Methods of determining whether a sequence is specifically hybridizable to a target nucleic acid are well known in the art. In one embodiment, the antisense compounds provided herein are specifically hybridizable with a PCSK9 nucleic acid.

The PCSK9 sequences targeted by some of the preferred antisense compounds disclosed in Table 1 are shown in FIG. 1. The antisense compounds are labeled with the corresponding Isis number. The bar in FIG. 1 labeled “NM_(—)174936.2” displays the relative position of the target sequences within GENBANK® Accession No. NM_(—)174936.2, first deposited with GENBANK® on Jun. 1, 2003 (SEQ ID NO: 1). The bar below that shows the corresponding structural elements of the PCSK9 mRNA, where “CDS” is the protein-encoding region of the mRNA and “UTRs” are the 5′ and 3′ untranslated regions. As shown in FIG. 1, the sequences targeted by the present antisense compounds are found in the protein-encoding region of the PCSK9 mRNA.

Complementarity

An antisense compound and a target nucleic acid are complementary to each other when a sufficient number of nucleobases of the antisense compound can hydrogen bond with the corresponding nucleobases of the target nucleic acid, such that a desired effect will occur (e.g., antisense inhibition of a target nucleic acid, such as a PCSK9 nucleic acid).

Non-complementary nucleobases between an antisense compound and a PCSK9 nucleic acid may be tolerated provided that the antisense compound remains able to specifically hybridize to a target nucleic acid. Moreover, an antisense compound may hybridize over one or more segments of a PCSK9 nucleic acid such that intervening or adjacent segments are not involved in the hybridization event (e.g., a loop structure, mismatch or hairpin structure).

In some embodiments, the antisense compounds provided herein are at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% complementary to a PCSK9 nucleic acid. Percent complementarity of an antisense compound with a target nucleic acid can be determined using routine methods.

In other embodiments, the antisense compounds provided herein are fully complementary (i.e, 100% complementary) to a target nucleic acid. For example, antisense compound may be fully complementary to a PCSK9 nucleic acid. As used herein, “fully complementary” means each nucleobase of an antisense compound is capable of precise base pairing with the corresponding nucleobases of a target nucleic acid.

The location of a non-complementary nucleobase may be at the 5′ end or 3′ end of the antisense compound. Alternatively, the non-complementary nucleobase or nucleobases may be at an internal position of the antisense compound. When two or more non-complementary nucleobases are present, they may be contiguous (i.e., linked) or non-contiguous. In one embodiment, a non-complementary nucleobase is located in the wing segment of a gapmer antisense oligonucleotide.

In one embodiment, antisense compounds up to 20 nucleobases in length comprise no more than 4, no more than 3, no more than 2 or no more than 1 non-complementary nucleobase(s) relative to a target nucleic acid, such as a PCSK9 nucleic acid.

In another embodiment, antisense compounds up to 30 nucleobases in length comprise no more than 6, no more than 5, no more than 4, no more than 3, no more than 2 or no more than 1 non-complementary nucleobase(s) relative to a target nucleic acid, such as a PCSK9 nucleic acid.

The antisense compounds provided herein also include those which are complementary to a portion of a target nucleic acid. As used herein, “portion” refers to a defined number of contiguous (i.e., linked) nucleobases within a region or segment of a target nucleic acid. A “portion” can also refer to a defined number of contiguous nucleobases of an antisense compound. In one embodiment, the antisense compounds are complementary to at least an 8 nucleobase portion of a target segment. In another embodiment, the antisense compounds are complementary to at least a 12 nucleobase portion of a target segment. In yet another embodiment, the antisense compounds are complementary to at least a 15 nucleobase portion of a target segment.

Identity

The antisense compounds provided herein may also have a defined percent identity to a particular nucleotide sequence, SEQ ID NO, or compound represented by a specific Isis number. As used herein, an antisense compound is identical to the sequence disclosed herein if it has the same nucleobase pairing ability. For example, a RNA which contains uracil in place of thymidine in a disclosed DNA sequence would be considered identical to the DNA sequence since both uracil and thymidine pair with adenine. Shortened and lengthened versions of the antisense compounds described herein as well as compounds having non-identical bases relative to the antisense compounds provided herein also are contemplated. The non-identical bases may be adjacent to each other or dispersed throughout the antisense compound. Percent identity of an antisense compound is calculated according to the number of bases that have identical base pairing relative to the sequence to which it is being compared.

In one embodiment, the antisense compounds are at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% identical to one or more of the antisense compounds disclosed herein.

Modifications

A nucleoside is a base-sugar combination. The nucleobase (also known as base) portion of the nucleoside is normally a heterocyclic base moiety. Nucleotides are nucleosides that further include a phosphate group covalently linked to the sugar portion of the nucleoside. For those nucleosides that include a pentofuranosyl sugar, the phosphate group can be linked to the 2′, 3′ or 5′ hydroxyl moiety of the sugar. Oligonucleotides are formed through the covalent linkage of adjacent nucleosides to one another, to form a linear polymeric oligonucleotide. Within the oligonucleotide structure, the phosphate groups are commonly referred to as forming the internucleoside linkages of the oligonucleotide.

Modifications to antisense compounds encompass substitutions or changes to internucleoside linkages, sugar moieties, or nucleobases. Modified antisense compounds are often preferred over native forms because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for nucleic acid target, increased stability in the presence of nucleases, or increased inhibitory activity.

Chemically modified nucleosides may also be employed to increase the binding affinity of a shortened or truncated antisense oligonucleotide for its target nucleic acid. Consequently, comparable results can often be obtained with shorter antisense compounds that have such chemically modified nucleosides.

Modified Internucleoside Linkages

The naturally occurring internucleoside linkage of RNA and DNA is a 3′ to 5′ phosphodiester linkage. Antisense compounds having one or more modified, i.e., non-naturally occurring, internucleoside linkages are often selected over antisense compounds having naturally occurring internucleoside linkages because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for target nucleic acids, and increased stability in the presence of nucleases.

Oligonucleotides having modified internucleoside linkages include internucleoside linkages that retain a phosphorus atom as well as internucleoside linkages that do not have a phosphorus atom. Representative phosphorus containing internucleoside linkages include, but are not limited to, phosphodiesters, phosphotriesters, methylphosphonates, phosphoramidate, and phosphorothioates. Methods of preparation of phosphorous-containing and non-phosphorous-containing linkages are well known.

In one embodiment, antisense compounds targeted to a PCSK9 nucleic acid comprise one or more modified internucleoside linkages. In some embodiments, the modified internucleoside linkages are phosphorothioate linkages. In other embodiments, each internucleoside linkage of an antisense compound is a phosphorothioate internucleoside linkage.

Modified Sugar Moieties

Antisense compounds targeted to a PCSK9 nucleic acid may contain one or more nucleotides having modified sugar moieties. Sugar modifications may impart nuclease stability, binding affinity or some other beneficial biological property to the antisense compounds. The furanosyl sugar ring of a nucleoside can be modified in a number of ways including, but not limited to: addition of a substituent group, particularly at the 2′ position; bridging of two non-geminal ring atoms to form a bicyclic nucleic acid (BNA); and substitution of an atom or group such as —S—, —N(R)— or —C(R₁)(R₂) (R═H, C1-C12 alkyl or a protecting group) for the ring oxygen at the 4′-position and combinations of these such as for example a 2′-F-5′-methyl substituted nucleoside (see WO 2008/101157, published Aug. 21, 2008, for other disclosed 5′,2′-bis substituted nucleosides) or replacement of the ribosyl ring oxygen atom with S with further substitution at the 2′-position (see U.S. Published Application No. 2005/0130923, published on Jun. 16, 2005) or alternatively 5′-substitution of a BNA (see WO 2007/134181, published Nov. 22, 2007, wherein BNA is substituted with for example a 5′-methyl or a 5′-vinyl group).

Modified sugars include, but are not limited to: substituted sugars, especially 2′-substituted sugars having a 2′-F, 2′-OCH₃ (2′-OMe) or a 2′-O(CH₂)₂—OCH₃ (2′-O-methoxyethyl or 2′-MOE) substituent group; and bicyclic modified sugars (BNAs), having a 4′-(CH₂)_(n)—O-2′ bridge, where n=1 or n=2, including α-L-methyleneoxy (4′-CH2-O-2′ or 4′-α-L-(CH2)-O-2′ (α-L-LNA)) BNA, β-D-methyleneoxy (4′-CH2-O-2′ or 4′-β-D-(CH2)-O-2′ (β-D-LNA)) BNA and ethyleneoxy (4′-(CH2)₂-O-2′ (ENA)) BNA. BNAs also include 4′-(CH2)-S-2′ and 4′-C(CH3)2-O-2′ (see International Application No. PCT/US2008/068922); 4′-CH(CH3)-O-2′ and 4′-CH(CH2OCH3)-O-2′ (see U.S. Pat. No. 7,399,845, issued Jul. 15, 2008); 4′-CH2-N(OCH3)-2′ (see International Application No. PCT/US2008/064591); 4′-CH2-O—N(CH3)-2′ (see U.S. Published Application No. 2004/0171570, published Sep. 2, 2004); 4′-CH2-N(R)—O-2′ (see U.S. Pat. No. 7,427,672, issued Sep. 23, 2008); 4′-CH2-C(CH3)-2′ and 4′-CH2-C(═CH2)-2′ (see International Application No. PCT/US2008/066154); and wherein R is, independently, H, C1-C12 alkyl, or a protecting group. Bicyclic modified sugars also include (6′S)-6′methyl BNA, aminooxy (4′-CH2-O—N(R)-2′) BNA, oxyamino (4′-CH2-N(R)—O-2′) BNA wherein R is, independently, H, a protecting group or C1-C12 alkyl. Examples of nucleosides having modified sugar moieties also include without limitation nucleosides comprising 5′-vinyl, 5′-methyl (R or S) and 4′-S.

The substituent at the 2′ position can also be selected from allyl, amino, azido, thio, O-allyl, O—C1-C10 alkyl, OCF₃, O(CH₂)₂SCH₃, O(CH₂)₂—O—N(Rm)(Rn), and O—CH₂—C(═O)—N(Rm)(Rn), where each Rm and Rn is, independently, H or substituted or unsubstituted C1-C10 alkyl.

In certain embodiments, such BNA modified nucleotides are high-affinity nucleotides and their incorporation into antisense compounds allows for increased potency and improved therapeutic index. Methods for the preparations of modified sugars are well known to those skilled in the art.

In certain embodiments, nucleosides are modified by substitution of the ribosyl ring with a sugar surrogate. Such modification includes without limitation, replacement of the ribosyl ring with a surrogate ring system (sometimes referred to as DNA analogs) such as a morpholino ring, a cyclohexenyl ring, a cyclohexyl ring or a tetrahydropyranyl ring such as one having one of the formula:

Many other bicyclo and tricyclo sugar surrogate ring systems are also know in the art that can be used to modify nucleosides for incorporation into antisense compounds. Such ring systems can undergo various additional substitutions to enhance activity.

Methods for the preparations of modified sugars are well known to those skilled in the art.

In nucleotides having modified sugar moieties (including substitution of a sugar surrogate), the nucleobase moieties (natural, modified or a combination thereof) are maintained for hybridization with an appropriate nucleic acid target.

In one embodiment, antisense compounds targeted to a PCSK9 nucleic acid comprise one or more nucleotides having modified sugar moieties. In a suitable embodiment, the modified sugar moiety is 2′-MOE. In other embodiments, the 2′-MOE modified nucleotides are arranged in a gapmer motif.

Modified Nucleobases

Nucleobase (or base) modifications or substitutions are structurally distinguishable from, yet functionally interchangeable with, naturally occurring or synthetic unmodified nucleobases. Both natural and modified nucleobases are capable of participating in hydrogen bonding. Such nucleobase modifications may impart nuclease stability, binding affinity or some other beneficial biological property to antisense compounds. Modified nucleobases include synthetic and natural nucleobases such as, for example, 5-methylcytosine (5-me-C). Certain nucleobase substitutions, including 5-methylcytosine substitutions, are particularly useful for increasing the binding affinity of an antisense compound for a target nucleic acid. For example, 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2° C. (Sanghvi, Y. S., Crooke, S. T. and Lebleu, B., eds., Antisense Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278).

Additional unmodified nucleobases include 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl (—C≡C—CH₃) uracil and cytosine and other alkynyl derivatives of pyrimidine bases, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 2-F-adenine, 2-amino-adenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-2.5 deazaguanine and 3-deazaadenine.

Heterocyclic base moieties may also include those in which the purine or pyrimidine base is replaced with other heterocycles, for example 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine and 2-pyridone. Nucleobases that are particularly useful for increasing the binding affinity of antisense compounds include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O-6 substituted purines, including 2 aminopropyladenine, 5-propynyluracil and 5-propynylcytosine.

In one embodiment, antisense compounds targeted to a PCSK9 nucleic acid comprise one or more modified nucleobases. In an additional embodiment, gap-widened antisense oligonucleotides targeted to a PCSK9 nucleic acid comprise one or more modified nucleobases. In some embodiments, the modified nucleobase is 5-methylcytosine. In further embodiments, each cytosine is a 5-methylcytosine.

Compositions and Methods for Formulating Pharmaceutical Compositions

Antisense oligonucleotides may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions are dependent upon a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

Antisense compound targeted to a PCSK9 nucleic acid can be utilized in pharmaceutical compositions by combining the antisense compound with a suitable pharmaceutically acceptable diluent or carrier. A pharmaceutically acceptable diluent includes phosphate-buffered saline (PBS). PBS is a diluent suitable for use in compositions to be delivered parenterally. Accordingly, in one embodiment, employed in the methods described herein is a pharmaceutical composition comprising an antisense compound targeted to a PCSK9 nucleic acid and a pharmaceutically acceptable diluent. In one embodiment, the pharmaceutically acceptable diluent is PBS. In other embodiments, the antisense compound is an antisense oligonucleotide.

Pharmaceutical compositions comprising antisense compounds encompass any pharmaceutically acceptable salts, esters, or salts of such esters, or any other oligonucleotide which, upon administration to an animal, including a human, is capable of providing (directly or indirectly) the biologically active metabolite or residue thereof. Accordingly, for example, the disclosure is also drawn to pharmaceutically acceptable salts of antisense compounds, prodrugs, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

A prodrug can include the incorporation of additional nucleosides at one or both ends of an antisense compound which are cleaved by endogenous nucleases within the body, to form the active antisense compound.

In certain embodiments, pharmaceutical compositions of the present invention comprise one or more oligonucleotides and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulosem and polyvinylpyrrolidone.

In certain embodiments, a pharmaceutical composition of the present invention is prepared using known techniques, including, but not limited to mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or tabletting processes.

In certain embodiments, a pharmaceutical composition of the present invention is a liquid (e.g., a suspension, elixir and/or solution). In certain of such embodiments, a liquid pharmaceutical composition is prepared using ingredients known in the art, including, but not limited to, water, glycols, oils, alcohols, flavoring agents, preservatives, and coloring agents.

In certain embodiments, a pharmaceutical composition of the present invention is a solid (e.g., a powder, tablet, and/or capsule). In certain of such embodiments, a solid pharmaceutical composition comprising one or more oligonucleotides is prepared using ingredients known in the art, including, but not limited to, starches, sugars, diluents, granulating agents, lubricants, binders, and disintegrating agents.

In certain embodiments, a pharmaceutical composition of the present invention is formulated as a depot preparation. Certain such depot preparations are typically longer acting than non-depot preparations. In certain embodiments, such preparations are administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. In certain embodiments, depot preparations are prepared using suitable polymeric or hydrophobic materials (for example an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

In certain embodiments, a pharmaceutical composition of the present invention comprises a delivery system. Examples of delivery systems include, but are not limited to, liposomes and emulsions. Certain delivery systems are useful for preparing certain pharmaceutical compositions including those comprising hydrophobic compounds. In certain embodiments, certain organic solvents such as dimethylsulfoxide are used.

In certain embodiments, a pharmaceutical composition of the present invention comprises one or more tissue-specific delivery molecules designed to deliver the one or more pharmaceutical agents of the present invention to specific tissues or cell types. For example, in certain embodiments, pharmaceutical compositions include liposomes coated with a tissue-specific antibody.

In certain embodiments, a pharmaceutical composition of the present invention comprises a co-solvent system. Certain of such co-solvent systems comprise, for example, benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. In certain embodiments, such co-solvent systems are used for hydrophobic compounds. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80™, and 65% w/v polyethylene glycol 300. The proportions of such co-solvent systems may be varied considerably without significantly altering their solubility and toxicity characteristics. Furthermore, the identity of co-solvent components may be varied: for example, other surfactants may be used instead of Polysorbate 80™; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose.

In certain embodiments, a pharmaceutical composition of the present invention comprises a sustained-release system. A non-limiting example of such a sustained-release system is a semi-permeable matrix of solid hydrophobic polymers. In certain embodiments, sustained-release systems may, depending on their chemical nature, release pharmaceutical agents over a period of hours, days, weeks or months.

In certain embodiments, a pharmaceutical composition of the present invention is prepared for oral administration. In certain of such embodiments, a pharmaceutical composition is formulated by combining one or more oligonucleotides with one or more pharmaceutically acceptable carriers. Certain of such carriers enable pharmaceutical compositions to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a subject. In certain embodiments, pharmaceutical compositions for oral use are obtained by mixing oligonucleotide and one or more solid excipient. Suitable excipients include, but are not limited to, fillers, such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). In certain embodiments, such a mixture is optionally ground and auxiliaries are optionally added. In certain embodiments, pharmaceutical compositions are formed to obtain tablets or dragee cores. In certain embodiments, disintegrating agents (e.g., cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate) are added.

In certain embodiments, dragee cores are provided with coatings. In certain such embodiments, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to tablets or dragee coatings.

In certain embodiments, pharmaceutical compositions for oral administration are push-fit capsules made of gelatin. Certain of such push-fit capsules comprise one or more pharmaceutical agents of the present invention in admixture with one or more filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In certain embodiments, pharmaceutical compositions for oral administration are soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. In certain soft capsules, one or more pharmaceutical agents of the present invention are be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added.

In certain embodiments, pharmaceutical compositions are prepared for buccal administration. Certain of such pharmaceutical compositions are tablets or lozenges formulated in conventional manner.

In certain embodiments, a pharmaceutical composition is prepared for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In certain of such embodiments, a pharmaceutical composition comprises a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In certain embodiments, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives). In certain embodiments, injectable suspensions are prepared using appropriate liquid carriers, suspending agents and the like. Certain pharmaceutical compositions for injection are presented in unit dosage form, e.g., in ampoules or in multi-dose containers. Certain pharmaceutical compositions for injection are suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Certain solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, such suspensions may also contain suitable stabilizers or agents that increase the solubility of the pharmaceutical agents to allow for the preparation of highly concentrated solutions.

In certain embodiments, a pharmaceutical composition is prepared for transmucosal administration. In certain of such embodiments penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

In certain embodiments, a pharmaceutical composition is prepared for administration by inhalation. Certain of such pharmaceutical compositions for inhalation are prepared in the form of an aerosol spray in a pressurized pack or a nebulizer. Certain of such pharmaceutical compositions comprise a propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In certain embodiments using a pressurized aerosol, the dosage unit may be determined with a valve that delivers a metered amount. In certain embodiments, capsules and cartridges for use in an inhaler or insufflator may be formulated. Certain of such formulations comprise a powder mixture of a pharmaceutical agent of the invention and a suitable powder base such as lactose or starch.

In certain embodiments, a pharmaceutical composition is prepared for rectal administration, such as a suppositories or retention enema. Certain of such pharmaceutical compositions comprise known ingredients, such as cocoa butter and/or other glycerides.

In certain embodiments, a pharmaceutical composition is prepared for topical administration. Certain of such pharmaceutical compositions comprise bland moisturizing bases, such as ointments or creams. Exemplary suitable ointment bases include, but are not limited to, petrolatum, petrolatum plus volatile silicones, lanolin and water in oil emulsions such as Eucerin™, available from Beiersdorf (Cincinnati, Ohio). Exemplary suitable cream bases include, but are not limited to, Nivea™ Cream, available from Beiersdorf (Cincinnati, Ohio), cold cream (USP), Purpose Cream™, available from Johnson & Johnson (New Brunswick, N.J.), hydrophilic ointment (USP) and Lubriderm™, available from Pfizer (Morris Plains, N.J.).

In certain embodiments, a pharmaceutical composition of the present invention comprises an oligonucleotide in a therapeutically effective amount. In certain embodiments, the therapeutically effective amount is sufficient to prevent, alleviate or ameliorate symptoms of a disease or to prolong the survival of the subject being treated. Determination of a therapeutically effective amount is well within the capability of those skilled in the art.

In certain embodiments, one or more oligonucleotides of the present invention is formulated as a prodrug. In certain embodiments, upon in vivo administration, a prodrug is chemically converted to the biologically, pharmaceutically or therapeutically more active form of the oligonucleotide. In certain embodiments, prodrugs are useful because they are easier to administer than the corresponding active form. For example, in certain instances, a prodrug may be more bioavailable (e.g., through oral administration) than is the corresponding active form. In certain instances, a prodrug may have improved solubility compared to the corresponding active form. In certain embodiments, prodrugs are less water soluble than the corresponding active form. In certain instances, such prodrugs possess superior transmittal across cell membranes, where water solubility is detrimental to mobility. In certain embodiments, a prodrug is an ester. In certain such embodiments, the ester is metabolically hydrolyzed to carboxylic acid upon administration. In certain instances the carboxylic acid containing compound is the corresponding active form. In certain embodiments, a prodrug comprises a short peptide (polyaminoacid) bound to an acid group. In certain of such embodiments, the peptide is cleaved upon administration to form the corresponding active form.

In certain embodiments, a prodrug is produced by modifying a pharmaceutically active compound such that the active compound will be regenerated upon in vivo administration. The prodrug can be designed to alter the metabolic stability or the transport characteristics of a drug, to mask side effects or toxicity, to improve the flavor of a drug or to alter other characteristics or properties of a drug. By virtue of knowledge of pharmacodynamic processes and drug metabolism in vivo, those of skill in this art, once a pharmaceutically active compound is known, can design prodrugs of the compound (see, e.g., Nogrady (1985) Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392).

In certain embodiments, a pharmaceutical composition comprising one or more pharmaceutical agents of the present invention is useful for treating a conditions or disorders in a mammalian, and particularly in a human, subject. Suitable administration routes include, but are not limited to, oral, rectal, transmucosal, intestinal, enteral, topical, suppository, through inhalation, intrathecal, intraventricular, intraperitoneal, intranasal, intraocular and parenteral (e.g., intravenous, intramuscular, intramedullary, and subcutaneous). In certain embodiments, pharmaceutical intrathecals are administered to achieve local rather than systemic exposures. For example, pharmaceutical compositions may be injected directly in the area of desired effect (e.g., in the renal or cardiac area).

In certain embodiments, a pharmaceutical composition of the present invention is administered in the form of a dosage unit (e.g., tablet, capsule, bolus, etc.). In certain embodiments, such pharmaceutical compositions comprise an oligonucleotide in a dose selected from 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg, 130 mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165 mg, 170 mg, 175 mg, 180 mg, 185 mg, 190 mg, 195 mg, 200 mg, 205 mg, 210 mg, 215 mg, 220 mg, 225 mg, 230 mg, 235 mg, 240 mg, 245 mg, 250 mg, 255 mg, 260 mg, 265 mg, 270 mg, 270 mg, 280 mg, 285 mg, 290 mg, 295 mg, 300 mg, 305 mg, 310 mg, 315 mg, 320 mg, 325 mg, 330 mg, 335 mg, 340 mg, 345 mg, 350 mg, 355 mg, 360 mg, 365 mg, 370 mg, 375 mg, 380 mg, 385 mg, 390 mg, 395 mg, 400 mg, 405 mg, 410 mg, 415 mg, 420 mg, 425 mg, 430 mg, 435 mg, 440 mg, 445 mg, 450 mg, 455 mg, 460 mg, 465 mg, 470 mg, 475 mg, 480 mg, 485 mg, 490 mg, 495 mg, 500 mg, 505 mg, 510 mg, 515 mg, 520 mg, 525 mg, 530 mg, 535 mg, 540 mg, 545 mg, 550 mg, 555 mg, 560 mg, 565 mg, 570 mg, 575 mg, 580 mg, 585 mg, 590 mg, 595 mg, 600 mg, 605 mg, 610 mg, 615 mg, 620 mg, 625 mg, 630 mg, 635 mg, 640 mg, 645 mg, 650 mg, 655 mg, 660 mg, 665 mg, 670 mg, 675 mg, 680 mg, 685 mg, 690 mg, 695 mg, 700 mg, 705 mg, 710 mg, 715 mg, 720 mg, 725 mg, 730 mg, 735 mg, 740 mg, 745 mg, 750 mg, 755 mg, 760 mg, 765 mg, 770 mg, 775 mg, 780 mg, 785 mg, 790 mg, 795 mg, and 800 mg. In certain such embodiments, a pharmaceutical composition of the present invention comprises a dose of oligonucleotide selected from 25 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 500 mg, 600 mg, 700 mg, and 800 mg. In certain embodiments, a pharmaceutical composition is comprises a dose of oligonucleotide selected from 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, and 400 mg. In certain embodiments the dose is administered at intervals ranging from more than once per day, once per day, once per week, twice per week, three times per week, four times per week, five times per week, 6 times per week, once per month to once per three months, for as long as needed to sustain the desired effect.

In a further aspect, a pharmaceutical agent is sterile lyophilized oligonucleotide that is reconstituted with a suitable diluent, e.g., sterile water for injection. The reconstituted product is administered as a subcutaneous injection or as an intravenous infusion after dilution into saline. The lyophilized drug product consists of the oligonucleotide which has been prepared in water for injection, adjusted to pH 7.0-9.0 with acid or base during preparation, and then lyophilized. The lyophilized oligonucleotide may be 25-800 mg of the oligonucleotide. It is understood that this encompasses 25, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 725, 750, 775, and 800 mg of lyophilized oligonucleotide. The lyophilized drug product may be packaged in a 2 mL Type I, clear glass vial (ammonium sulfate-treated), stoppered with a bromobutyl rubber closure and sealed with an aluminum FLIP-OFF® overseal. In one embodiment, the lyophilized pharmaceutical agent comprises ISIS 405879.

The compositions of the present invention may additionally contain other adjunct components conventionally found in pharmaceutical compositions, at their art-established usage levels. Thus, for example, the compositions may contain additional, compatible, pharmaceutically-active materials such as, for example, antipruritics, astringents, local anesthetics or anti-inflammatory agents, or may contain additional materials useful in physically formulating various dosage forms of the compositions of the present invention, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers. However, such materials, when added, should not unduly interfere with the biological activities of the components of the compositions of the present invention. The formulations can be sterilized and, if desired, mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings and/or aromatic substances and the like which do not deleteriously interact with the oligonucleotide(s) of the formulation.

Conjugated Antisense Compounds

Antisense compounds may be covalently linked to one or more moieties or conjugates which enhance the activity, visualization of cellular distribution, or cellular uptake of the resulting antisense oligonucleotides. Typical conjugate groups include cholesterol moieties and lipid moieties. Additional conjugate groups include carbohydrates, phospholipids, biotin, phenazine, folate, phenanthridine, anthraquinone, acridine, fluoresceins, rhodamines, coumarins, and dyes.

Antisense compounds can also be modified to have one or more stabilizing groups that are generally attached to one or both termini of antisense compounds to enhance properties such as, for example, nuclease stability. Included in stabilizing groups are cap structures. These terminal modifications protect the antisense compound having terminal nucleic acid from exonuclease degradation, and can help in delivery and/or localization within a cell. The cap can be present at the 5′-terminus (5′-cap), or at the 3′-terminus (3′-cap), or can be present on both termini. Cap structures are well known in the art and include, for example, inverted deoxy abasic caps. Further 3′ and 5′-stabilizing groups that can be used to cap one or both ends of an antisense compound to impart nuclease stability include those disclosed in WO 03/004602 published on Jan. 16, 2003.

Cell Culture and Antisense Compounds Treatment

The effects of antisense compounds on the level, activity or expression of PCSK9 nucleic acids can be tested in vitro in a variety of cell types. Cell types used for such analyses are available from commercial vendors (e.g., American Type Culture Collection, Manassus, Va.; Zen-Bio, Inc., Research Triangle Park, N.C.; Clonetics Corporation, Walkersville, Md.) and cells are cultured according to the vendor's instructions using commercially available reagents (e.g., Invitrogen Life Technologies, Carlsbad, Calif.). Illustrative cell types include, but are not limited to, HepG2 cells, Hep3B cells, and primary hepatocytes.

In Vitro Testing of Antisense Oligonucleotides

Described herein are methods for treatment of cells with antisense oligonucleotides, which can be modified appropriately for treatment with other antisense compounds.

In general, cells are treated with antisense oligonucleotides when they cells reach approximately 60-80% confluency in culture.

One reagent commonly used to introduce antisense oligonucleotides into cultured cells includes the cationic lipid transfection reagent LIPOFECTIN® (Invitrogen, Carlsbad, Calif.). Antisense oligonucleotides are mixed with LIPOFECTIN® in OPTI-MEM® 1 (Invitrogen, Carlsbad, Calif.) to achieve the desired final concentration of antisense oligonucleotide and a LIPOFECTIN® concentration that typically ranges 2 to 12 ug/mL per 100 nM antisense oligonucleotide.

Another reagent used to introduce antisense oligonucleotides into cultured cells includes LIPOFECTAMINE® (Invitrogen, Carlsbad, Calif.). Antisense oligonucleotide is mixed with LIPOFECTAMINE® in OPTI-MEM® 1 reduced serum medium (Invitrogen, Carlsbad, Calif.) to achieve the desired concentration of antisense oligonucleotide and a LIPOFECTAMINE® concentration that typically ranges 2 to 12 ug/mL per 100 nM antisense oligonucleotide.

Cells are treated with antisense oligonucleotides by routine methods. Cells are typically harvested 16-24 hours after antisense oligonucleotide treatment, at which time RNA or protein levels of target nucleic acids are measured by methods known in the art and described herein. In general, when treatments are performed in multiple replicates, the data are presented as the average of the replicate treatments.

The concentration of antisense oligonucleotide used varies from cell line to cell line. Methods to determine the optimal antisense oligonucleotide concentration for a particular cell line are well known in the art. Antisense oligonucleotides are typically used at concentrations ranging from 1 nM to 300 mM.

RNA Isolation

RNA analysis can be performed on total cellular RNA or poly(A)+ mRNA. Methods of RNA isolation are well known in the art. RNA is prepared using methods well known in the art, for example, using the TRIZOL® Reagent (Invitrogen, Carlsbad, Calif.) according to the manufacturer's recommended protocols.

Analysis of Inhibition of Target Levels or Expression

Inhibition of levels or expression of a PCSK9 nucleic acid can be assayed in a variety of ways known in the art. For example, target nucleic acid levels can be quantitated by, e.g., Northern blot analysis, competitive polymerase chain reaction (PCR), or quantitative real-time PCR. RNA analysis can be performed on total cellular RNA or poly(A)+ mRNA. Methods of RNA isolation are well known in the art. Northern blot analysis is also routine in the art. Quantitative real-time PCR can be conveniently accomplished using the commercially available ABI PRISM® 7600, 7700, or 7900 Sequence Detection System, available from PE-Applied Biosystems, Foster City, Calif. and used according to manufacturer's instructions.

Quantitative Real-Time PCR Analysis of Target RNA Levels

Quantitation of target RNA levels may be accomplished by quantitative real-time PCR using the ABI PRISMS 7600, 7700, or 7900 Sequence Detection System (PE-Applied Biosystems, Foster City, Calif.) according to manufacturer's instructions. Methods of quantitative real-time PCR are well known in the art.

Prior to real-time PCR, the isolated RNA is subjected to a reverse transcriptase (RT) reaction, which produces complementary DNA (cDNA) that is then used as the substrate for the real-time PCR amplification. The RT and real-time PCR reactions are performed sequentially in the same sample well. RT and real-time PCR reagents are obtained from Invitrogen (Carlsbad, Calif.). RT, real-time-PCR reactions are carried out by methods well known to those skilled in the art.

Gene (or RNA) target quantities obtained by real time PCR are normalized using either the expression level of a gene whose expression is constant, such as GAPDH, or by quantifying total RNA using RIBOGREEN® (Invitrogen, Inc. Carlsbad, Calif.). GAPDH expression is quantified by real time PCR, by being run simultaneously with the target, multiplexing, or separately. Total RNA is quantified using RIBOGREEN® RNA quantification reagent (Invitrogen, Inc. Carlsbad, Calif.). Methods of RNA quantification by RIBOGREEN® are taught in Jones, L. J., et al, (Analytical Biochemistry, 1998, 265, 368-374). A CYTOFLUOR® 4000 instrument (PE Applied Biosystems) is used to measure RIBOGREEN® fluorescence.

Probes and primers are designed to hybridize to a PCSK9 nucleic acid. Methods for designing real-time PCR probes and primers are well known in the art, and may include the use of software such as PRIMER EXPRESS® Software (Applied Biosystems, Foster City, Calif.).

Analysis of Protein Levels

Antisense inhibition of PCSK9 nucleic acids can be assessed by measuring PCSK9 protein levels. Protein levels of PCSK9 can be evaluated or quantitated in a variety of ways well known in the art, such as immunoprecipitation, Western blot analysis (immunoblotting), enzyme-linked immunosorbent assay (ELISA), quantitative protein assays, protein activity assays (for example, caspase activity assays), immunohistochemistry, immunocytochemistry or fluorescence-activated cell sorting (FACS). Antibodies directed to a target can be identified and obtained from a variety of sources, such as the MSRS catalog of antibodies (Aerie Corporation, Birmingham, Mich.), or can be prepared via conventional monoclonal or polyclonal antibody generation methods well known in the art. Antibodies useful for the detection of human and rat PCSK9 are commercially available.

Serum Lipid Analysis

Plasma concentrations of total cholesterol, LDL-C, HDL-C, free cholesterol, triglycerides, glucose, ketones, transaminases, and phospholipids can be measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, N.Y.). Serum lipoprotein and cholesterol profiling can be performed as described by Crooke et al., Lipid Res. (2005) 46: 872-884, using a Beckman System Gold 126 HPLC system, a 507e refrigerated autosampler, a 126 photodiode array detector (Beckman Instruments, Fullerton, Calif.), and a Superose 6 HR 10/30 column (Pfizer, Chicago, Ill.). HDL, LDL and VLDL fractions are measured at a wavelength of 505 nm and validated with a cholesterol calibration kit. (Sigma).

Liver Triglyceride Analysis

Liver triglyceride content can be measured according to routine experimental procedures, for example, per procedures described by Desai et al., Diabetes (2001) 50: 2287-2295. Liver triglyceride content may be measured after antisense inhibition of PCSK9 for 6 weeks.

In Vivo Testing of Antisense Compounds

Antisense compounds, for example, antisense oligonucleotides, are tested in animals to assess their ability to inhibit expression of PCSK9 and produce phenotypic changes, such as decreases in LDL-C. Testing may be performed in normal animals, or in experimental disease models. For administration to animals, antisense oligonucleotides are formulated in a pharmaceutically acceptable diluent, such as phosphate-buffered saline. Administration includes parenteral routes of administration, such as intraperitoneal, intravenous, and subcutaneous. Calculation of antisense oligonucleotide dosage and dosing frequency is within the abilities of those skilled in the art, and depends upon factors such as route of administration and animal body weight. Following a period of treatment with antisense oligonucleotides, RNA is isolated from various tissues and changes in PCSK9 nucleic acid expression are measured. Changes in PCSK9 protein levels may also be measured.

Nonlimiting Disclosure and Incorporation by Reference

The following examples serve only to illustrate the methods, antisense oligonucleotides, and compositions provided and are not intended to limit the same. Each of the references, GENBANK® accession numbers, and the like recited in the present application is incorporated herein by reference in its entirety.

EXAMPLES Example 1 Antisense Inhibition of Human PCSK9: A549 Cells

Antisense oligonucleotides targeted to a PCSK9 nucleic acid were tested for their effects on PCSK9 mRNA in vitro. Cultured A549 cells at a density of 4000 cells per well in a 96-well plate were treated with 60 nM of antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. PCSK9 mRNA levels were adjusted according to total RNA content as measured by RIBOGREEN®. Results are presented as percent inhibition of PCSK9, relative to untreated control cells. Antisense oligonucleotides that exhibited at least 30% inhibition of PCSK9 expression are shown in Table 16.

The motif column indicates the wing-gap-wing motif of each antisense oligonucleotide. Antisense oligonucleotides were designed as 5-10-5 gapmers, or alternatively as 3-14-3 gapmers, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises 2′-MOE nucleotides. As illustrated in Table 16, a single nucleobase sequence may be represented by a 5-10-5 motif as well as a 3-14-3 motif. “5′ target site” indicates the 5′-most nucleotide which the antisense oligonucleotide is targeted on the indicated GENBANK Accession No.

TABLE 16 Antisense inhibition of PCSK9 in human cells (Cell type A549) SEQ 5′ ID Target Target Nucleic % IsisNo NO Site Acid Sequence Motif Inhibition 395150 5 242 NM_174936.2 GAGGAGACCTAGAGGCCGTG 5-10-5 57 395151 6 300 NM_174936.2 AGGACCGCCTGGAGCTGACG 5-10-5 77 395152 7 410 NM_174936.2 ACGCAAGGCTAGCACCAGCT 5-10-5 81 395153 9 480 NM_174936.2 CCTTGGCGCAGCGGTGGAAG 5-10-5 51 395154 10 561 NM_174936.2 GGCGGGCAGTGCGCTCTGAC 5-10-5 40 395155 11 600 NM_174936.2 TGGTGAGGTATCCCCGGCGG 5-10-5 79 395156 14 620 NM_174936.2 ATGGAAGACATGCAGGATCT 5-10-5 75 395157 15 646 NM_174936.2 TTCACCAGGAAGCCAGGAAG 5-10-5 72 395158 17 705 NM_174936.2 CCTCGATGTAGTCGACATGG 5-10-5 61 395159 18 785 NM_174936.2 GTATTCATCCGCCCGGTACC 5-10-5 35 395160 19 835 NM_174936.2 CTGGTGTCTAGGAGATACAC 5-10-5 47 395161 21 860 NM_174936.2 GATTTCCCGGTGGTCACTCT 5-10-5 70 395162 24 890 NM_174936.2 CTCGAAGTCGGTGACCATGA 5-10-5 52 395163 25 923 NM_174936.2 GTGGAAGCGGGTCCCGTCCT 5-10-5 73 395164 26 970 NM_174936.2 ACCCCTGCCAGGTGGGTGCC 5-10-5 69 395165 28 1004 NM_174936.2 ACCCTTGGCCACGCCGGCAT 5-10-5 81 395166 29 1040 NM_174936.2 TTGGCAGTTGAGCACGCGCA 5-10-5 65 395167 31 1077 NM_174936.2 CCAGGCCTATGAGGGTGCCG 5-10-5 34 395168 32 1098 NM_174936.2 GCTGGCTTTTCCGAATAAAC 5-10-5 69 395169 33 1210 NM_174936.2 GTGACCAGCACGACCCCAGC 5-10-5 61 395171 34 1326 NM_174936.2 CCAAAGTCCCCAGGGTCACC 5-10-5 38 395173 36 1340 NM_174936.2 GCCAAAGTTGGTCCCCAAAG 5-10-5 64 395174 38 1361 NM_174936.2 GGCAAAGAGGTCCACACAGC 5-10-5 72 395175 39 1389 NM_174936.2 TGGAGGCACCAATGATGTCC 5-10-5 41 395177 45 1534 NM_174936.2 TTGGCAGAGAAGTGGATCAG 5-10-5 41 395178 50 1569 NM_174936.2 GGTCCTCAGGGAACCAGGCC 5-10-5 67 395181 52 1640 NM_174936.2 AAACAGCTGCCAACCTGCCC 5-10-5 37 395182 54 1675 NM_174936.2 GTAGGCCCCGAGTGTGCTGA 5-10-5 51 395183 57 1812 NM_174936.2 CGTTGTGGGCCCGGCAGACC 5-10-5 74 395184 58 1858 NM_174936.2 AGCAGGCAGCACCTGGCAAT 5-10-5 61 395185 59 1920 NM_174936.2 CACGGGTCCCCATGCTGGCC 5-10-5 73 395186 60 2100 NM_174936.2 CTTTGCATTCCAGACCTGGG 5-10-5 83 395187 62 2310 NM_174936.2 GGCAGCAGATGGCAACGGCT 5-10-5 80 395189 64 2504 NM_174936.2 TCAAGGGCCAGGCCAGCAGC 5-10-5 65 395190 66 2597 NM_174936.2 ATGCCCCACAGTGAGGGAGG 5-10-5 60 395191 67 2606 NM_174936.2 AATGGTGAAATGCCCCACAG 5-10-5 76 395193 68 2750 NM_174936.2 CATGGGAAGAATCCTGCCTC 5-10-5 48 395194 69 2832 NM_174936.2 GGAGATGAGGGCCATCAGCA 5-10-5 64 395195 70 2900 NM_174936.2 TAGATGCCATCCAGAAAGCT 5-10-5 44 395196 72 2983 NM_174936.2 GGCATAGAGCAGAGTAAAGG 5-10-5 39 395198 75 3437 NM_174936.2 GCTCAAGGAGGGACAGTTGT 5-10-5 73 395199 76 3472 NM_174936.2 AAAGATAAATGTCTGCTTGC 5-10-5 83 395200 78 3543 NM_174936.2 TCTTCAAGTTACAAAAGCAA 5-10-5 40 395202 129 13816 NT_032977.8 GTGCCATCTGAACAGCACCT 5-10-5 36 395203 110 13926 NT_032977.8 CCTGGAACCCCTGCAGCCAG 5-10-5 64 395204 152 13977 NT_032977.8 TTCAGGCAGGTTGCTGCTAG 5-10-5 55 395206 136 14122 NT_032977.8 TAGGAGAAAGTAGGGAGAGC 5-10-5 45 395207 132 14179 NT_032977.8 TAAAAGCTGCAAGAGACTCA 5-10-5 49 395208 139 14267 NT_032977.8 TCAGAGAAAACAGTCACCGA 5-10-5 56 395209 142 14404 NT_032977.8 TCATTTTAGAGACAGGAAGC 5-10-5 73 395210 113 14441 NT_032977.8 GAATAACAGTGATGTCTGGC 5-10-5 82 395211 138 14494 NT_032977.8 TCACAGCTCACCGAGTCTGC 5-10-5 60 395212 98 14524 NT_032977.8 AGTGTAAAATAAAGCCCCTA 5-10-5 50 395213 96 14601 NT_032977.8 AGGACCCAAGTCATCCTGCT 5-10-5 57 395214 124 14631 NT_032977.8 GGCCATCAGCTGGCAATGCT 5-10-5 50 395215 133 14675 NT_032977.8 TAGACAAGGAAAGGGAGGCC 5-10-5 60 395216 103 14681 NT_032977.8 ATTTCATAGACAAGGAAAGG 5-10-5 51 395218 146 1315 AK124635.1 TGACATTTGTGGGAGAGGAG 5-10-5 35 395220 104 2085 AK124635.1 CACAGTCCTGCAAAACAGCT 5-10-5 54 395221 102 5590 NT_032977.8 ATGTGCAGAGATCAATCACA 5-10-5 69 395222 127 10633 NT_032977.8 GGTGGTAATTTGTCACAGCA 5-10-5 64 395223 84 11308 NT_032977.8 AAGGTCACACAGTTAAGAGT 5-10-5 55 395226 147 24858 NT_032977.8 TGAGTTCATTTAAGAGTGGA 5-10-5 42 399793 8 417 NM_174936.2 CCTCGGAACGCAAGGCTAGC 5-10-5 71 399794 12 606 NM_174936.2 GGATCTTGGTGAGGTATCCC 5-10-5 42 399795 13 615 NM_174936.2 AGACATGCAGGATCTTGGTG 5-10-5 54 399796 16 651 NM_174936.2 TCATCTTCACCAGGAAGCCA 5-10-5 50 399797 20 840 NM_174936.2 GTATGCTGGTGTCTAGGAGA 5-10-5 38 399798 22 866 NM_174936.2 GCCCTCGATTTCCCGGTGGT 5-10-5 54 399799 23 880 NM_174936.2 GTGACCATGACCCTGCCCTC 5-10-5 75 399800 27 975 NM_174936.2 TGACCACCCCTGCCAGGTGG 5-10-5 67 399801 30 1045 NM_174936.2 TTCCCTTGGCAGTTGAGCAC 5-10-5 79 399802 35 1335 NM_174936.2 AGTTGGTCCCCAAAGTCCCC 5-10-5 71 399803 37 1352 NM_174936.2 GTCCACACAGCGGCCAAAGT 5-10-5 63 399804 40 1400 NM_174936.2 GCTGCAGTCGCTGGAGGCAC 5-10-5 46 399805 41 1411 NM_174936.2 ACAAAGCAGGTGCTGCAGTC 5-10-5 46 399806 42 1470 NM_174936.2 GCATCATGGCTGCAATGCCA 5-10-5 85 399807 43 1478 NM_174936.2 GGCAGACAGCATCATGGCTG 5-10-5 71 399808 44 1526 NM_174936.2 GAAGTGGATCAGTCTCTGCC 5-10-5 38 399809 46 1539 NM_174936.2 CATCTTTGGCAGAGAAGTGG 5-10-5 53 399810 47 1545 NM_174936.2 TGATGACATCTTTGGCAGAG 5-10-5 63 399811 48 1552 NM_174936.2 GCCTCATTGATGACATCTTT 5-10-5 61 399812 49 1564 NM_174936.2 TCAGGGAACCAGGCCTCATT 5-10-5 46 399813 51 1583 NM_174936.2 GGTCAGTACCCGCTGGTCCT 5-10-5 72 399814 53 1645 NM_174936.2 CTGCAAAACAGCTGCCAACC 5-10-5 88 399815 55 1740 NM_174936.2 AGAAACTGGAGCAGCTCAGC 5-10-5 38 399816 56 1746 NM_174936.2 TCCTGGAGAAACTGGAGCAG 5-10-5 51 399817 61 2105 NM_174936.2 CTTGACTTTGCATTCCAGAC 5-10-5 84 399818 63 2415 NM_174936.2 AACCATTTTAAAGCTCAGCC 5-10-5 68 399819 65 2509 NM_174936.2 CCCACTCAAGGGCCAGGCCA 5-10-5 87 399821 71 2906 NM_174936.2 TCTGGCTAGATGCCATCCAG 5-10-5 83 399822 73 2988 NM_174936.2 AGCCTGGCATAGAGCAGAGT 5-10-5 70 399824 74 3233 NM_174936.2 AAGTAAGAAGAGGCTTGGCT 5-10-5 43 399825 77 3477 NM_174936.2 ACCCAAAAGATAAATGTCTG 5-10-5 61 399827 100 13746 NT_032977.8 ATCTCAGGACAGGTGAGCAA 5-10-5 81 399828 116 13760 NT_032977.8 GAGTAGAGATTCTCATCTCA 5-10-5 73 399829 117 13828 NT_032977.8 GAGTCTTCTGAAGTGCCATC 5-10-5 65 399831 83 13986 NT_032977.8 AAGGAAGACTTCAGGCAGGT 5-10-5 52 399833 92 14397 NT_032977.8 AGAGACAGGAAGCTGCAGCT 5-10-5 54 399834 82 14670 NT_032977.8 AAGGAAAGGGAGGCCTAGAG 5-10-5 44 399836 130 2095 AK124635.1 GTGCTGACCACACAGTCCTG 5-10-5 96 399837 107 3056 NT_032977.8 CCCACTATAATGGCAAGCCC 5-10-5 83 399838 80 4306 NT_032977.8 AACCCAGTTCTAATGCACCT 5-10-5 81 399839 106 5140 NT_032977.8 CCAGTCAGAGTAGAACAGAG 5-10-5 68 399840 121 5599 NT_032977.8 GGAGCCTACATGTGCAGAGA 5-10-5 46 399841 94 5667 NT_032977.8 AGCATGGCACCAGCATCTGC 5-10-5 59 399842 108 6652 NT_032977.8 CCCAGCCCTATCAGGAAGTG 5-10-5 67 399843 144 7099 NT_032977.8 TGACATCCAGGAGGGAGGAG 5-10-5 33 399844 91 7556 NT_032977.8 AGACTGATGGAAGGCATTGA 5-10-5 56 399845 131 7565 NT_032977.8 GTGTTGAGCAGACTGATGGA 5-10-5 48 399846 145 8836 NT_032977.8 TGACATCTTGTCTGGGAGCC 5-10-5 58 399847 90 8948 NT_032977.8 AGACTAGGAGCCTGAGTTTT 5-10-5 38 399849 148 10252 NT_032977.8 TGGCAGCAACTCAGACATAT 5-10-5 74 399851 88 12715 NT_032977.8 ACTGGATACATTGGCAGACA 5-10-5 62 399852 111 12928 NT_032977.8 CTAGAGGAACCACTAGATAT 5-10-5 66 399854 97 16134 NT_032977.8 AGTCAAGCTGCTGCCCAGAG 5-10-5 82 399855 120 16668 NT_032977.8 GCTAGTTATTAAGCACCTGC 5-10-5 71 399856 150 17267 NT_032977.8 TGTGAGCTCTGGCCCAGTGG 5-10-5 64 399857 115 18377 NT_032977.8 GAGTAAGGCAGGTTACTCTC 5-10-5 79 399858 134 18408 NT_032977.8 TAGATGTGACTAACATTTAA 5-10-5 57 399859 105 19203 NT_032977.8 CACATTAGCCTTGCTCAAGT 5-10-5 75 399860 151 19913 NT_032977.8 TGTGATGACCTGGAAAGGTG 5-10-5 36 399862 109 20188 NT_032977.8 CCCCTGCACAGAGCCTGGCA 5-10-5 59 399863 141 20624 NT_032977.8 TCATGGCTGCAATGCCTGGT 5-10-5 46 399864 93 20995 NT_032977.8 AGAGAGGAGGGCTTAAAGAA 5-10-5 30 399865 95 21082 NT_032977.8 AGCTGCCAACCTGCAAAAAG 5-10-5 75 399866 143 21481 NT_032977.8 TGAAAATCCATCCAGCACTG 5-10-5 74 399867 89 21589 NT_032977.8 AGAACCATGGAGCACCTGAG 5-10-5 84 399868 123 21696 NT_032977.8 GGCACTGCCCTTCCACCAAA 5-10-5 53 399869 118 24907 NT_032977.8 GCACCATCCAGACCAGAATC 5-10-5 49 399870 114 25413 NT_032977.8 GAGAGGTTCAGATCCAGGCC 5-10-5 66 399871 4 135 NM_174936.2 GCGCGGAATCCTGGCTGGGA 3-14-3 40 399872 5 242 NM_174936.2 GAGGAGACCTAGAGGCCGTG 3-14-3 64 399873 6 300 NM_174936.2 AGGACCGCCTGGAGCTGACG 3-14-3 65 399874 7 410 NM_174936.2 ACGCAAGGCTAGCACCAGCT 3-14-3 80 399875 9 480 NM_174936.2 CCTTGGCGCAGCGGTGGAAG 3-14-3 67 399876 10 561 NM_174936.2 GGCGGGCAGTGCGCTCTGAC 3-14-3 62 399877 11 600 NM_174936.2 TGGTGAGGTATCCCCGGCGG 3-14-3 73 399878 14 620 NM_174936.2 ATGGAAGACATGCAGGATCT 3-14-3 54 399879 15 646 NM_174936.2 TTCACCAGGAAGCCAGGAAG 3-14-3 75 399880 17 705 NM_174936.2 CCTCGATGTAGTCGACATGG 3-14-3 80 399881 18 785 NM_174936.2 GTATTCATCCGCCCGGTACC 3-14-3 63 399882 19 835 NM_174936.2 CTGGTGTCTAGGAGATACAC 3-14-3 66 399883 21 860 NM_174936.2 GATTTCCCGGTGGTCACTCT 3-14-3 59 399884 24 890 NM_174936.2 CTCGAAGTCGGTGACCATGA 3-14-3 71 399885 25 923 NM_174936.2 GTGGAAGCGGGTCCCGTCCT 3-14-3 73 399886 26 970 NM_174936.2 ACCCCTGCCAGGTGGGTGCC 3-14-3 76 399887 28 1004 NM_174936.2 ACCCTTGGCCACGCCGGCAT 3-14-3 54 399888 29 1040 NM_174936.2 TTGGCAGTTGAGCACGCGCA 3-14-3 75 399889 31 1077 NM_174936.2 CCAGGCCTATGAGGGTGCCG 3-14-3 84 399890 32 1098 NM_174936.2 GCTGGCTTTTCCGAATAAAC 3-14-3 73 399891 33 1210 NM_174936.2 GTGACCAGCACGACCCCAGC 3-14-3 99 399892 149 1297 NM_174936.2 TGGGCATTGGTGGCCCCAAC 3-14-3 61 399893 34 1326 NM_174936.2 CCAAAGTCCCCAGGGTCACC 3-14-3 54 399894 128 1330 NM_174936.2 GTCCCCAAAGTCCCCAGGGT 3-14-3 80 399895 36 1340 NM_174936.2 GCCAAAGTTGGTCCCCAAAG 3-14-3 82 399896 38 1361 NM_174936.2 GGCAAAGAGGTCCACACAGC 3-14-3 83 399897 39 1389 NM_174936.2 TGGAGGCACCAATGATGTCC 3-14-3 70 399898 101 1465 NM_174936.2 ATGGCTGCAATGCCAGCCAC 3-14-3 37 399899 45 1534 NM_174936.2 TTGGCAGAGAAGTGGATCAG 3-14-3 64 399900 50 1569 NM_174936.2 GGTCCTCAGGGAACCAGGCC 3-14-3 82 399901 87 1576 NM_174936.2 ACCCGCTGGTCCTCAGGGAA 3-14-3 76 399902 119 1605 NM_174936.2 GCAGGGCGGCCACCAGGTTG 3-14-3 39 399903 52 1640 NM_174936.2 AAACAGCTGCCAACCTGCCC 3-14-3 73 399904 54 1675 NM_174936.2 GTAGGCCCCGAGTGTGCTGA 3-14-3 81 399905 57 1812 NM_174936.2 CGTTGTGGGCCCGGCAGACC 3-14-3 80 399906 58 1858 NM_174936.2 AGCAGGCAGCACCTGGCAAT 3-14-3 92 399907 59 1920 NM_174936.2 CACGGGTCCCCATGCTGGCC 3-14-3 95 399908 60 2100 NM_174936.2 CTTTGCATTCCAGACCTGGG 3-14-3 84 399909 62 2310 NM_174936.2 GGCAGCAGATGGCAACGGCT 3-14-3 73 399910 154 2410 NM_174936.2 TTTTAAAGCTCAGCCCCAGC 3-14-3 57 399911 64 2504 NM_174936.2 TCAAGGGCCAGGCCAGCAGC 3-14-3 82 399912 66 2597 NM_174936.2 ATGCCCCACAGTGAGGGAGG 3-14-3 68 399913 67 2606 NM_174936.2 AATGGTGAAATGCCCCACAG 3-14-3 92 399914 153 2649 NM_174936.2 TTGGGAGCAGCTGGCAGCAC 3-14-3 59 399915 68 2750 NM_174936.2 CATGGGAAGAATCCTGCCTC 3-14-3 78 399916 69 2832 NM_174936.2 GGAGATGAGGGCCATCAGCA 3-14-3 71 399917 70 2900 NM_174936.2 TAGATGCCATCCAGAAAGCT 3-14-3 81 399918 72 2983 NM_174936.2 GGCATAGAGCAGAGTAAAGG 3-14-3 82 399919 112 3227 NM_174936.2 GAAGAGGCTTGGCTTCAGAG 3-14-3 57 399920 75 3437 NM_174936.2 GCTCAAGGAGGGACAGTTGT 3-14-3 84 399921 76 3472 NM_174936.2 AAAGATAAATGTCTGCTTGC 3-14-3 78 399922 78 3543 NM_174936.2 TCTTCAAGTTACAAAAGCAA 3-14-3 61 399923 85 13681 NT_032977.8 ACAAATTCCCAGACTCAGCA 3-14-3 82 399924 129 13816 NT_032977.8 GTGCCATCTGAACAGCACCT 3-14-3 84 399925 110 13926 NT_032977.8 CCTGGAACCCCTGCAGCCAG 3-14-3 73 399926 152 13977 NT_032977.8 TTCAGGCAGGTTGCTGCTAG 3-14-3 47 399927 140 13998 NT_032977.8 TCAGCCAGGCCAAAGGAAGA 3-14-3 78 399928 136 14122 NT_032977.8 TAGGAGAAAGTAGGGAGAGC 3-14-3 47 399929 132 14179 NT_032977.8 TAAAAGCTGCAAGAGACTCA 3-14-3 69 399930 139 14267 NT_032977.8 TCAGAGAAAACAGTCACCGA 3-14-3 82 399931 142 14404 NT_032977.8 TCATTTTAGAGACAGGAAGC 3-14-3 79 399932 113 14441 NT_032977.8 GAATAACAGTGATGTCTGGC 3-14-3 83 399933 138 14494 NT_032977.8 TCACAGCTCACCGAGTCTGC 3-14-3 78 399934 98 14524 NT_032977.8 AGTGTAAAATAAAGCCCCTA 3-14-3 79 399935 96 14601 NT_032977.8 AGGACCCAAGTCATCCTGCT 3-14-3 89 399936 124 14631 NT_032977.8 GGCCATCAGCTGGCAATGCT 3-14-3 70 399937 133 14675 NT_032977.8 TAGACAAGGAAAGGGAGGCC 3-14-3 78 399938 103 14681 NT_032977.8 ATTTCATAGACAAGGAAAGG 3-14-3 67 399940 146 1315 AK124635.1 TGACATTTGTGGGAGAGGAG 3-14-3 35 399942 104 2085 AK124635.1 CACAGTCCTGCAAAACAGCT 3-14-3 75 399943 102 5590 NT_032977.8 ATGTGCAGAGATCAATCACA 3-14-3 64 399944 127 10633 NT_032977.8 GGTGGTAATTTGTCACAGCA 3-14-3 73 399945 84 11308 NT_032977.8 AAGGTCACACAGTTAAGAGT 3-14-3 89 399947 126 22292 NT_032977.8 GGTGCATAAGGAGAAAGAGA 3-14-3 70 399948 147 24858 NT_032977.8 TGAGTTCATTTAAGAGTGGA 3-14-3 46 399949 8 417 NM_174936.2 CCTCGGAACGCAAGGCTAGC 3-14-3 74 399950 12 606 NM_174936.2 GGATCTTGGTGAGGTATCCC 3-14-3 68 399951 13 615 NM_174936.2 AGACATGCAGGATCTTGGTG 3-14-3 67 399952 16 651 NM_174936.2 TCATCTTCACCAGGAAGCCA 3-14-3 74 399953 20 840 NM_174936.2 GTATGCTGGTGTCTAGGAGA 3-14-3 59 399954 22 866 NM_174936.2 GCCCTCGATTTCCCGGTGGT 3-14-3 83 399955 23 880 NM_174936.2 GTGACCATGACCCTGCCCTC 3-14-3 62 399956 27 975 NM_174936.2 TGACCACCCCTGCCAGGTGG 3-14-3 78 399957 30 1045 NM_174936.2 TTCCCTTGGCAGTTGAGCAC 3-14-3 82 399958 35 1335 NM_174936.2 AGTTGGTCCCCAAAGTCCCC 3-14-3 78 399959 37 1352 NM_174936.2 GTCCACACAGCGGCCAAAGT 3-14-3 89 399960 40 1400 NM_174936.2 GCTGCAGTCGCTGGAGGCAC 3-14-3 64 399961 41 1411 NM_174936.2 ACAAAGCAGGTGCTGCAGTC 3-14-3 63 399962 42 1470 NM_174936.2 GCATCATGGCTGCAATGCCA 3-14-3 68 399963 43 1478 NM_174936.2 GGCAGACAGCATCATGGCTG 3-14-3 70 399964 44 1526 NM_174936.2 GAAGTGGATCAGTCTCTGCC 3-14-3 70 399965 46 1539 NM_174936.2 CATCTTTGGCAGAGAAGTGG 3-14-3 49 399966 47 1545 NM_174936.2 TGATGACATCTTTGGCAGAG 3-14-3 78 399967 48 1552 NM_174936.2 GCCTCATTGATGACATCTTT 3-14-3 82 399968 49 1564 NM_174936.2 TCAGGGAACCAGGCCTCATT 3-14-3 71 399969 51 1583 NM_174936.2 GGTCAGTACCCGCTGGTCCT 3-14-3 82 399970 53 1645 NM_174936.2 CTGCAAAACAGCTGCCAACC 3-14-3 74 399971 55 1740 NM_174936.2 AGAAACTGGAGCAGCTCAGC 3-14-3 48 399972 56 1746 NM_174936.2 TCCTGGAGAAACTGGAGCAG 3-14-3 63 399973 61 2105 NM_174936.2 CTTGACTTTGCATTCCAGAC 3-14-3 74 399974 63 2415 NM_174936.2 AACCATTTTAAAGCTCAGCC 3-14-3 71 399975 65 2509 NM_174936.2 CCCACTCAAGGGCCAGGCCA 3-14-3 91 399976 122 2582 NM_174936.2 GGAGGGAGCTTCCTGGCACC 3-14-3 67 399977 71 2906 NM_174936.2 TCTGGCTAGATGCCATCCAG 3-14-3 89 399978 73 2988 NM_174936.2 AGCCTGGCATAGAGCAGAGT 3-14-3 80 399979 135 2994 NM_174936.2 TAGCACAGCCTGGCATAGAG 3-14-3 83 399980 74 3233 NM_174936.2 AAGTAAGAAGAGGCTTGGCT 3-14-3 50 399981 77 3477 NM_174936.2 ACCCAAAAGATAAATGTCTG 3-14-3 55 399982 99 3550 NM_174936.2 ATAAATATCTTCAAGTTACA 3-14-3 36 399983 100 13746 NT_032977.8 ATCTCAGGACAGGTGAGCAA 3-14-3 96 399984 116 13760 NT_032977.8 GAGTAGAGATTCTCATCTCA 3-14-3 71 399985 117 13828 NT_032977.8 GAGTCTTCTGAAGTGCCATC 3-14-3 76 399986 81 13903 NT_032977.8 AAGCAGGGCCTCAGGTGGAA 3-14-3 68 399987 83 13986 NT_032977.8 AAGGAAGACTTCAGGCAGGT 3-14-3 59 399988 137 14112 NT_032977.8 TAGGGAGAGCTCACAGATGC 3-14-3 31 399989 92 14397 NT_032977.8 AGAGACAGGAAGCTGCAGCT 3-14-3 65 399990 82 14670 NT_032977.8 AAGGAAAGGGAGGCCTAGAG 3-14-3 59 399992 130 2095 AK124635.1 GTGCTGACCACACAGTCCTG 3-14-3 94 399993 107 3056 NT_032977.8 CCCACTATAATGGCAAGCCC 3-14-3 76 399994 80 4306 NT_032977.8 AACCCAGTTCTAATGCACCT 3-14-3 82 399995 106 5140 NT_032977.8 CCAGTCAGAGTAGAACAGAG 3-14-3 74 399996 121 5599 NT_032977.8 GGAGGCTACATGTGCAGAGA 3-14-3 65 399997 94 5667 NT_032977.8 AGCATGGCACCAGCATCTGC 3-14-3 77 399998 108 6652 NT_032977.8 CCCAGCCCTATCAGGAAGTG 3-14-3 78 399999 144 7099 NT_032977.8 TGACATCCAGGAGGGAGGAG 3-14-3 43 400000 91 7556 NT_032977.8 AGACTGATGGAAGGCATTGA 3-14-3 84 400001 131 7565 NT_032977.8 GTGTTGAGCAGACTGATGGA 3-14-3 73 400002 145 8836 NT_032977.8 TGACATCTTGTCTGGGAGCC 3-14-3 82 400003 90 8948 NT_032977.8 AGACTAGGAGCCTGAGTTTT 3-14-3 48 400004 125 9099 NT_032977.8 GGCCTGCAGAAGCCAGAGAG 3-14-3 45 400005 148 10252 NT_032977.8 TGGCAGCAACTCAGACATAT 3-14-3 73 400006 79 11472 NT_032977.8 AAATGCAGGGCTAAAATCAC 3-14-3 78 400007 88 12715 NT_032977.8 ACTGGATACATTGGCAGACA 3-14-3 77 400008 111 12928 NT_032977.8 CTAGAGGAACCACTAGATAT 3-14-3 78 400009 86 15471 NT_032977.8 ACAGCATTCTTGGTTAGGAG 3-14-3 73 400010 97 16134 NT_032977.8 AGTCAAGCTGCTGCCCAGAG 3-14-3 74 400011 120 16668 NT_032977.8 GCTAGTTATTAAGCACCTGC 3-14-3 75 400012 150 17267 NT_032977.8 TGTGAGCTCTGGCCCAGTGG 3-14-3 64 400013 115 18377 NT_032977.8 GAGTAAGGCAGGTTACTCTC 3-14-3 88 400014 134 18408 NT_032977.8 TAGATGTGACTAACATTTAA 3-14-3 58 400015 105 19203 NT_032977.8 CACATTAGCCTTGCTCAAGT 3-14-3 89 400018 109 20188 NT_032977.8 CCCCTGCACAGAGCCTGGCA 3-14-3 62 400019 141 20624 NT_032977.8 TCATGGCTGCAATGCCTGGT 3-14-3 47 400020 93 20995 NT_032977.8 AGAGAGGAGGGCTTAAAGAA 3-14-3 66 400021 95 21082 NT_032977.8 AGCTGCCAACCTGCAAAAAG 3-14-3 61 400022 143 21481 NT_032977.8 TGAAAATCCATCCAGCACTG 3-14-3 90 400023 89 21589 NT_032977.8 AGAACCATGGAGCACCTGAG 3-14-3 83 400024 123 21696 NT_032977.8 GGCACTGCCCTTCCACCAAA 3-14-3 79 400025 118 24907 NT_032977.8 GCACCATCCAGACCAGAATC 3-14-3 78 400026 114 25413 NT_032977.8 GAGAGGTTCAGATCCAGGCC 3-14-3 81

Antisense oligonucleotides with the following ISIS Nos exhibited at least 80% inhibition of PCSK9 mRNA levels: 399806, 399814, 399817, 399819, 399821, 399827, 399836, 399837, 399854, 399867, 399889, 399891, 399906, 399907, 399908, 399913, 399920, 399924, 399935, 399945, 399959, 399975, 399977, 399983, 399992, 400000, 400013, 400015, 400022, and 400023. The target segments to which these antisense oligonucleotides are targeted are active target segments. The target regions to which these antisense oligonucleotides are targeted are active target regions.

ISIS Nos 399814, 399819, 399836, 399891, 399906, 399907, 399913, 399935, 399945, 399959, 399975, 399977, 399983, 399992, 400013, 400015, and 400022 each exhibited at least 85% inhibition of PCSK9 mRNA levels.

ISIS Nos 399836, 399891, 399906, 399907, 399913, 399975, 399983, 399992, and 400022 each exhibited at least 90% inhibition of PCSK9 mRNA levels.

Example 2 Antisense Inhibition of Human PCSK9 in HepG2 Cells

Antisense oligonucleotides were tested for their ability to inhibit the expression of PCSK9 mRNA in cultured HepG2 cells. HepG2 cells at a density of 10000 cells per well in a 96-well plate were treated with 1500 nM of antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. PCSK9 mRNA levels were adjusted according to total RNA content as measured by RIBOGREEN®. Results are presented as percent inhibition of PCSK9, relative to untreated control cells.

Isis Nos. 399819, 395149, 395150, 395151, 395152, 395153, 395154, 395155, 395156, 395157, 395158, 395161, 395162, 395163, 395164, 395165, 395166, 395167, 395168, 395172, 395173, 395174, 395178, 395179, 395182, 395183, 395185, 395186, 395187, 395189, 395190, 395191, 395193, 395194, 395195, 395198, 395199, 395203, 395207, 395210, 395211, 395213, 395214, 395221, 395222, 399793, 399794, 399795, 399798, 399801, 399803, 399804, 399806, 399807, 399812, 399813, 399814, 399816, 399817, 399820, 399821, 399822, 399823, 399827, 399828, 399829, 399833, 399834, 399836, 399837, 399841, 399844, 399846, 399848, 399849, 399851, 399852, 399854, 399855, 399856, 399857, 399862, 399865, 399867, 399868, 399872, 399873, 399874, 399875, 399876, 399877, 399879, 399880, 399883, 399884, 399885, 399886, 399887, 399888, 399889, 399890, 399891, 399892, 399894, 399895, 399896, 399900, 399901, 399902, 399904, 399905, 399906, 399907, 399908, 399909, 399911, 399912, 399913, 399915, 399916, 399918, 399920, 399921, 399924, 399925, 399929, 399931, 399935, 399936, 399937, 399943, 399944, 399945, 399949, 399950, 399954, 399957, 399959, 399960, 399962, 399963, 399966, 399967, 399968, 399969, 399970, 399972, 399973, 399975, 399976, 399977, 399978, 399979, 399983, 399984, 399985, 399986, 399989, 399990, 399992, 399996, 399997, 399998, 400000, 400001, 400002, 400004, 400005, 400007, 400008, 400009, 400010, 400012, 400013, 400015, 400018, 400021, 400023, 400024, 400025, and 400026 each inhibited PCSK9 by at least 70% in this experiment. The target segments to which these antisense oligonucleotides are targeted are active target segments. The target regions to which these antisense oligonucleotides are targeted are active target regions.

Isis Nos 399819, 395150, 395152, 395153, 395154, 395155, 395156, 395158, 395163, 395164, 395165, 395166, 395167, 395168, 395173, 395178, 395183, 395185, 395186, 395187, 395189, 395190, 395193, 395194, 395199, 395203, 395210, 395213, 395214, 399793, 399798, 399801, 399804, 399806, 399813, 399816, 399820, 399821, 399822, 399836, 399849, 399856, 399857, 399862, 399868, 399875, 399877, 399880, 399885, 399886, 399887, 399888, 399889, 399890, 399894, 399896, 399900, 399901, 399904, 399905, 399906, 399907, 399911, 399912, 399916, 399920, 399925, 399935, 399936, 399937, 399945, 399954, 399957, 399959, 399960, 399962, 399968, 399969, 399973, 399976, 399977, 399978, 399989, 399992, 400002, 400004, 400007, 400012, 400013, 400023, and 400024 each inhibited PCSK9 by at least 80% in this experiment.

Isis Nos 399819, 395152, 395153, 395155, 395156, 395158, 395163, 395164, 395165, 395166, 395168, 395178, 395183, 395185, 395187, 395189, 395190, 395194, 395199, 395203, 395214, 399793, 399804, 399813, 399821, 399822, 399836, 399856, 399868, 399877, 399887, 399888, 399890, 399900, 399904, 399905, 399907, 399911, 399912, 399916, 399925, 399936, 399954, 399960, 399962, 399969, 399976, 399977, 399978, 399989, 399992, 400012, 400013, 400023, and 400024 each inhibited PCSK9 by at least 85% in this experiment.

Isis Nos 399819, 395165, 395183, 395185, 399868, 399900, 399907, 399911, 399912, 399916, 399936, 399969, and 400024 each inhibited PCSK9 by at least 90% in this experiment.

Example 3 Antisense Inhibition of Human PCSK9 (Hep3B Cells)

Antisense oligonucleotides targeted to a PCSK9 nucleic acid were tested for their effects on PCSK9 mRNA in vitro. Cultured Hep3B cells at a density of 4000 cells per well in a 96-well plate were treated with 75 nM of antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. PCSK9 mRNA levels were adjusted according to total RNA content as measured by RIBOGREEN®. Results are presented as percent inhibition of PCSK9, relative to untreated control cells. Antisense oligonucleotides that exhibited at least 30% inhibition of PCSK9 expression are shown in Tables 17, 18, and 19.

The motif column indicates the wing-gap-wing motif of each antisense oligonucleotide. Antisense oligonucleotides were designed as 5-10-5 gapmers, or alternatively as 3-14-3 gapmers, or alternatively as 2-13-5 gapmers where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises 2′-MOE nucleotides. As illustrated in Table 17, 18 and 19, a single nucleobase sequence may be represented by a 5-10-5 motif as well as a 3-14-3 motif as well as a 2-13-5 motif. “5′ target site” indicates the 5′-most nucleotide which the antisense oligonucleotide is targeted on the indicated GENBANK Accession No.

TABLE 17 Antisense inhibition of PCSK9 in human cells (Hep3B) targeting SEQ ID NO: 1 5′ Target 3′ Target Site to Site to SEQ SEQ ID SEQ ID % ID Isis No. NO: 1 NO: 1 Sequence 5′-3′ Motif inhibition NO 410742 294 313 GCCTGGAGCTGACGGTGCCC 5-10-5 75.1 159 405999 298 317 GACCGCCTGGAGCTGACGGT 5-10-5 70.3 160 395151 300 319 AGGACCGCCTGGAGCTGACG 5-10-5 57.3 6 405861 406 425 AAGGCTAGCACCAGCTCCTC 5-10-5 90.5 162 405862 407 426 CAAGGCTAGCACCAGCTCCT 5-10-5 92.3 163 405863 408 427 GCAAGGCTAGCACCAGCTCC 5-10-5 82.6 164 405864 409 428 CGCAAGGCTAGCACCAGCTC 5-10-5 91.8 165 395152 410 429 ACGCAAGGCTAGCACCAGCT 5-10-5 93.3 7 405865 411 430 AACGCAAGGCTAGCACCAGC 5-10-5 84.6 166 405866 412 431 GAACGCAAGGCTAGCACCAG 5-10-5 81.8 167 405867 413 432 GGAACGCAAGGCTAGCACCA 5-10-5 74.0 168 405868 414 433 CGGAACGCAAGGCTAGCACC 5-10-5 76.8 169 399793 417 436 CCTCGGAACGCAAGGCTAGC 5-10-5 77.9 8 410743 421 440 TCCTCCTCGGAACGCAAGGC 5-10-5 76.7 170 410744 446 465 GTGCTCGGGTGCTTCGGCCA 5-10-5 76.9 171 410745 466 485 TGGAAGGTGGCTGTGGTTCC 5-10-5 41.7 172 395153 480 499 CCTTGGCGCAGCGGTGGAAG 5-10-5 81.1 9 406000 482 501 ATCCTTGGCGCAGCGGTGGA 5-10-5 79.1 173 406001 484 503 GGATCCTTGGCGCAGCGGTG 5-10-5 63.1 174 406002 488 507 CCACGGATCCTTGGCGCAGC 5-10-5 80.8 175 410746 507 526 CGTAGGTGCCAGGCAACCTC 5-10-5 44.8 176 410747 545 564 TGACTGCGAGAGGTGGGTCT 5-10-5 NA 177 406003 555 574 CAGTGCGCTCTGACTGCGAG 5-10-5 84.2 178 406004 557 576 GGCAGTGCGCTCTGACTGCG 5-10-5 56.4 179 406005 559 578 CGGGCAGTGCGCTCTGACTG 5-10-5 83.4 180 406006 562 581 CGGCGGGCAGTGCGCTCTGA 5-10-5 71.0 181 410748 591 610 ATCCCCGGCGGGCAGCCTGG 5-10-5 56.0 182 406007 595 614 AGGTATCCCCGGCGGGCAGC 5-10-5 76.3 183 410574 597 616 TGAGGTATCCCCGGCGGGCA 3-14-3 69.1 184 410529 597 616 TGAGGTATCCCCGGCGGGCA 5-10-5 77.4 184 410647 597 616 TGAGGTATCCCCGGCGGGCA 2-13-5 90.7 184 410575 598 617 GTGAGGTATCCCCGGCGGGC 3-14-3 71.5 185 410648 598 617 GTGAGGTATCCCCGGCGGGC 2-13-5 82.8 185 410530 598 617 GTGAGGTATCCCCGGCGGGC 5-10-5 83.2 185 410649 599 618 GGTGAGGTATCCCCGGCGGG 2-13-5 53.5 186 410576 599 618 GGTGAGGTATCCCCGGCGGG 3-14-3 56.2 186 410531 599 618 GGTGAGGTATCCCCGGCGGG 5-10-5 63.6 186 410650 600 619 TGGTGAGGTATCCCCGGCGG 2-13-5 86.1 11 399877 600 619 TGGTGAGGTATCCCCGGCGG 3-14-3 92.6 11 395155 600 619 TGGTGAGGTATCCCCGGCGG 5-10-5 92.7 11 410577 601 620 TTGGTGAGGTATCCCCGGCG 3-14-3 62.4 187 410532 601 620 TTGGTGAGGTATCCCCGGCG 5-10-5 63.5 187 410651 601 620 TTGGTGAGGTATCCCCGGCG 2-13-5 75.5 187 410652 602 621 CTTGGTGAGGTATCCCCGGC 2-13-5 81.2 188 410578 602 621 CTTGGTGAGGTATCCCCGGC 3-14-3 83.3 188 406008 602 621 CTTGGTGAGGTATCCCCGGC 5-10-5 95.2 188 410653 603 622 TCTTGGTGAGGTATCCCCGG 2-13-5 68.9 189 410579 603 622 TCTTGGTGAGGTATCCCCGG 3-14-3 74.0 189 410533 603 622 TCTTGGTGAGGTATCCCCGG 5-10-5 79.0 189 410580 604 623 ATCTTGGTGAGGTATCCCCG 3-14-3 62.3 190 410654 604 623 ATCTTGGTGAGGTATCCCCG 2-13-5 65.2 190 406009 604 623 ATCTTGGTGAGGTATCCCCG 5-10-5 90.7 190 410534 605 624 GATCTTGGTGAGGTATCCCC 5-10-5 59.8 191 410581 605 624 GATCTTGGTGAGGTATCCCC 3-14-3 72.9 191 410655 605 624 GATCTTGGTGAGGTATCCCC 2-13-5 77.4 191 399794 606 625 GGATCTTGGTGAGGTATCCC 5-10-5 56.5 12 410656 606 625 GGATCTTGGTGAGGTATCCC 2-13-5 68.0 12 399950 606 625 GGATCTTGGTGAGGTATCCC 3-14-3 74.4 12 410535 607 626 AGGATCTTGGTGAGGTATCC 5-10-5 65.7 192 410582 607 626 AGGATCTTGGTGAGGTATCC 3-14-3 69.5 192 410657 607 626 AGGATCTTGGTGAGGTATCC 2-13-5 73.1 192 406010 609 628 GCAGGATCTTGGTGAGGTAT 5-10-5 56.9 193 406011 611 630 ATGCAGGATCTTGGTGAGGT 5-10-5 70.7 194 406012 613 632 ACATGCAGGATCTTGGTGAG 5-10-5 69.1 195 406013 617 636 GAAGACATGCAGGATCTTGG 5-10-5 77.4 196 395156 620 639 ATGGAAGACATGCAGGATCT 5-10-5 76.7 14 410749 628 647 AGAAGGCCATGGAAGACATG 5-10-5 59.4 197 410750 638 657 GAAGCCAGGAAGAAGGCCAT 5-10-5 57.5 198 399879 646 665 TTCACCAGGAAGCCAGGAAG 3-14-3 91.9 15 406014 648 667 TCTTCACCAGGAAGCCAGGA 5-10-5 94.0 199 406015 653 672 ACTCATCTTCACCAGGAAGC 5-10-5 78.8 200 406016 655 674 CCACTCATCTTCACCAGGAA 5-10-5 87.2 201 410730 657 676 CGCCACTCATCTTCACCAGG 5-10-5 85.4 202 406017 659 678 GTCGCCACTCATCTTCACCA 5-10-5 76.3 203 406018 661 680 AGGTCGCCACTCATCTTCAC 5-10-5 63.2 204 406019 663 682 GCAGGTCGCCACTCATCTTC 5-10-5 68.1 205 406020 665 684 CAGCAGGTCGCCACTCATCT 5-10-5 83.7 206 410731 667 686 TCCAGCAGGTCGCCACTCAT 5-10-5 72.9 207 410751 685 704 GGCAACTTCAAGGCCAGCTC 5-10-5 84.6 208 395158 705 724 CCTCGATGTAGTCGACATGG 5-10-5 93.9 17 410752 724 743 GCAAAGACAGAGGAGTCCTC 5-10-5 76.3 209 406021 782 801 TTCATCCGCCCGGTACCGTG 5-10-5 79.6 210 406022 784 803 TATTCATCCGCCCGGTACCG 5-10-5 78.7 211 406023 787 806 TGGTATTCATCCGCCCGGTA 5-10-5 92.5 212 406024 789 808 GCTGGTATTCATCCGCCCGG 5-10-5 69.7 213 410732 791 810 GGGCTGGTATTCATCCGCCC 5-10-5 30.4 214 410753 821 840 ATACACCTCCACCAGGCTGC 5-10-5 72.8 215 406025 832 851 GTGTCTAGGAGATACACCTC 5-10-5 74.2 216 406026 837 856 TGCTGGTGTCTAGGAGATAC 5-10-5 80.5 217 405869 862 881 TCGATTTCCCGGTGGTCACT 5-10-5 87.6 218 405870 863 882 CTCGATTTCCCGGTGGTCAC 5-10-5 83.3 219 405871 864 883 CCTCGATTTCCCGGTGGTCA 5-10-5 87.0 220 405872 865 884 CCCTCGATTTCCCGGTGGTC 5-10-5 88.2 221 399798 866 885 GCCCTCGATTTCCCGGTGGT 5-10-5 84.3 22 399954 866 885 GCCCTCGATTTCCCGGTGGT 3-14-3 90.3 22 405873 867 886 TGCCCTCGATTTCCCGGTGG 5-10-5 90.0 222 405874 868 887 CTGCCCTCGATTTCCCGGTG 5-10-5 91.4 223 405875 869 888 CCTGCCCTCGATTTCCCGGT 5-10-5 93.5 224 405876 870 889 CCCTGCCCTCGATTTCCCGG 5-10-5 90.1 225 406027 874 893 ATGACCCTGCCCTCGATTTC 5-10-5 73.9 226 406028 876 895 CCATGACCCTGCCCTCGATT 5-10-5 92.3 227 406029 878 897 GACCATGACCCTGCCCTCGA 5-10-5 91.4 228 406030 882 901 CGGTGACCATGACCCTGCCC 5-10-5 95.6 229 406031 884 903 GTCGGTGACCATGACCCTGC 5-10-5 89.6 230 410733 886 905 AAGTCGGTGACCATGACCCT 5-10-5 65.7 231 406032 888 907 CGAAGTCGGTGACCATGACC 5-10-5 88.5 232 410754 898 917 GGCACATTCTCGAAGTCGGT 5-10-5 80.1 233 395163 923 942 GTGGAAGCGGGTCCCGTCCT 5-10-5 83.9 25 410755 933 952 TGGCCTGTCTGTGGAAGCGG 5-10-5 NA 234 410756 960 979 GGTGGGTGCCATGACTGTCA 5-10-5 67.1 235 406033 967 986 CCTGCCAGGTGGGTGCCATG 5-10-5 91.2 237 406034 972 991 CCACCCCTGCCAGGTGGGTG 5-10-5 59.3 238 406035 977 996 GCTGACCACCCCTGCCAGGT 5-10-5 91.7 239 410757 985 1004 TCCCGGCCGCTGACCACCCC 5-10-5 88.9 240 406036 989 1008 GGCATCCCGGCCGCTGACCA 5-10-5 84.0 241 406037 992 1011 GCCGGCATCCCGGCCGCTGA 5-10-5 55.9 242 410536 997 1016 GCCACGCCGGCATCCCGGCC 5-10-5 63.7 243 410658 997 1016 GCCACGCCGGCATCCCGGCC 2-13-5 82.0 243 410583 997 1016 GCCACGCCGGCATCCCGGCC 3-14-3 87.3 243 410537 998 1017 GGCCACGCCGGCATCCCGGC 5-10-5 58.5 244 410659 998 1017 GGCCACGCCGGCATCCCGGC 2-13-5 73.7 244 410584 998 1017 GGCCACGCCGGCATCCCGGC 3-14-3 79.5 244 410660 999 1018 TGGCCACGCCGGCATCCCGG 2-13-5 63.0 245 410585 999 1018 TGGCCACGCCGGCATCCCGG 3-14-3 73.2 245 406038 999 1018 TGGCCACGCCGGCATCCCGG 5-10-5 86.3 245 410661 1000 1019 TTGGCCACGCCGGCATCCCG 2-13-5 53.3 246 410586 1000 1019 TTGGCCACGCCGGCATCCCG 3-14-3 60.3 246 405877 1000 1019 TTGGCCACGCCGGCATCCCG 5-10-5 83.2 246 410587 1001 1020 CTTGGCCACGCCGGCATCCC 3-14-3 63.2 247 410662 1001 1020 CTTGGCCACGCCGGCATCCC 2-13-5 67.3 247 405878 1001 1020 CTTGGCCACGCCGGCATCCC 5-10-5 91.4 247 410588 1002 1021 CCTTGGCCACGCCGGCATCC 3-14-3 65.3 248 410663 1002 1021 CCTTGGCCACGCCGGCATCC 2-13-5 67.9 248 405879 1002 1021 CCTTGGCCACGCCGGCATCC 5-10-5 94.1 248 410589 1003 1022 CCCTTGGCCACGCCGGCATC 3-14-3 80.1 249 410664 1003 1022 CCCTTGGCCACGCCGGCATC 2-13-5 81.9 249 405880 1003 1022 CCCTTGGCCACGCCGGCATC 5-10-5 93.5 249 410665 1004 1023 ACCCTTGGCCACGCCGGCAT 2-13-5 78.7 28 399887 1004 1023 ACCCTTGGCCACGCCGGCAT 3-14-3 91.9 28 395165 1004 1023 ACCCTTGGCCACGCCGGCAT 5-10-5 96.7 28 410590 1005 1024 CACCCTTGGCCACGCCGGCA 3-14-3 82.2 250 410666 1005 1024 CACCCTTGGCCACGCCGGCA 2-13-5 82.7 250 405881 1005 1024 CACCCTTGGCCACGCCGGCA 5-10-5 98.3 250 410667 1006 1025 GCACCCTTGGCCACGCCGGC 2-13-5 84.7 251 410591 1006 1025 GCACCCTTGGCCACGCCGGC 3-14-3 86.3 251 405882 1006 1025 GCACCCTTGGCCACGCCGGC 5-10-5 91.6 251 410668 1007 1026 GGCACCCTTGGCCACGCCGG 2-13-5 91.4 252 405883 1007 1026 GGCACCCTTGGCCACGCCGG 5-10-5 92.5 252 410592 1007 1026 GGCACCCTTGGCCACGCCGG 3-14-3 93.4 252 410669 1008 1027 TGGCACCCTTGGCCACGCCG 2-13-5 69.0 253 410593 1008 1027 TGGCACCCTTGGCCACGCCG 3-14-3 88.8 253 405884 1008 1027 TGGCACCCTTGGCCACGCCG 5-10-5 90.8 253 410670 1009 1028 CTGGCACCCTTGGCCACGCC 2-13-5 74.9 254 410594 1009 1028 CTGGCACCCTTGGCCACGCC 3-14-3 75.2 254 410538 1009 1028 CTGGCACCCTTGGCCACGCC 5-10-5 78.2 254 410539 1010 1029 GCTGGCACCCTTGGCCACGC 5-10-5 61.9 255 410595 1010 1029 GCTGGCACCCTTGGCCACGC 3-14-3 83.1 255 410671 1010 1029 GCTGGCACCCTTGGCCACGC 2-13-5 83.3 255 410758 1015 1034 CGCATGCTGGCACCCTTGGC 5-10-5 73.6 256 406039 1036 1055 CAGTTGAGCACGCGCAGGCT 5-10-5 91.2 257 406040 1038 1057 GGCAGTTGAGCACGCGCAGG 5-10-5 96.5 258 399888 1040 1059 TTGGCAGTTGAGCACGCGCA 3-14-3 90.9 29 395166 1040 1059 TTGGCAGTTGAGCACGCGCA 5-10-5 96.2 29 406041 1042 1061 CCTTGGCAGTTGAGCACGCG 5-10-5 96.6 259 399801 1045 1064 TTCCCTTGGCAGTTGAGCAC 5-10-5 95.6 30 406042 1047 1066 CCTTCCCTTGGCAGTTGAGC 5-10-5 92.0 260 406043 1051 1070 GTGCCCTTCCCTTGGCAGTT 5-10-5 91.9 261 406044 1053 1072 CCGTGCCCTTCCCTTGGCAG 5-10-5 95.4 262 410759 1064 1083 GGTGCCGCTAACCGTGCCCT 5-10-5 83.7 263 410760 1088 1107 CCGAATAAACTCCAGGCCTA 5-10-5 96.7 265 406045 1096 1115 TGGCTTTTCCGAATAAACTC 5-10-5 88.4 266 395168 1098 1117 GCTGGCTTTTCCGAATAAAC 5-10-5 91.9 32 405909 1100 1119 CAGCTGGCTTTTCCGAATAA 5-10-5 80.3 267 405910 1102 1121 ACCAGCTGGCTTTTCCGAAT 5-10-5 90.9 268 405911 1104 1123 GGACCAGCTGGCTTTTCCGA 5-10-5 88.0 269 405912 1108 1127 GGCTGGACCAGCTGGCTTTT 5-10-5 78.2 270 410761 1119 1138 GTGGCCCCACAGGCTGGACC 5-10-5 53.7 271 410762 1132 1151 AGCAGCACCACCAGTGGCCC 5-10-5 57.6 272 410763 1154 1173 GCTGTACCCACCCGCCAGGG 5-10-5 68.8 273 410764 1200 1219 CGACCCCAGCCCTCGCCAGG 5-10-5 66.3 274 399891 1210 1229 GTGACCAGCACGACCCCAGC 3-14-3 91.1 33 405913 1212 1231 CGGTGACCAGCACGACCCCA 5-10-5 93.8 275 405914 1214 1233 AGCGGTGACCAGCACGACCC 5-10-5 90.4 276 405915 1216 1235 GCAGCGGTGACCAGCACGAC 5-10-5 86.8 277 405916 1218 1237 CGGCAGCGGTGACCAGCACG 5-10-5 91.3 278 410734 1219 1238 CCGGCAGCGGTGACCAGCAC 5-10-5 65.8 279 405917 1222 1241 TTGCCGGCAGCGGTGACCAG 5-10-5 62.0 280 405918 1224 1243 AGTTGCCGGCAGCGGTGACC 5-10-5 33.5 281 405919 1226 1245 GAAGTTGCCGGCAGCGGTGA 5-10-5 68.2 282 405920 1228 1247 CGGAAGTTGCCGGCAGCGGT 5-10-5 86.2 283 405921 1230 1249 CCCGGAAGTTGCCGGCAGCG 5-10-5 85.5 284 405922 1232 1251 GTCCCGGAAGTTGCCGGCAG 5-10-5 86.0 285 410765 1273 1292 ATGACCTCGGGAGCTGAGGC 5-10-5 50.4 286 410766 1283 1302 CCCAACTGTGATGACCTCGG 5-10-5 70.6 287 405923 1295 1314 GGCATTGGTGGCCCCAACTG 5-10-5 94.3 288 410767 1305 1324 GCTGGTCTTGGGCATTGGTG 5-10-5 65.1 289 405924 1318 1337 CCCAGGGTCACCGGCTGGTC 5-10-5 93.1 290 410735 1320 1339 TCCCCAGGGTCACCGGCTGG 5-10-5 78.1 291 405925 1322 1341 AGTCCCCAGGGTCACCGGCT 5-10-5 93.5 292 405926 1324 1343 AAAGTCCCCAGGGTCACCGG 5-10-5 94.6 293 405927 1328 1347 CCCCAAAGTCCCCAGGGTCA 5-10-5 94.5 294 405928 1333 1352 TTGGTCCCCAAAGTCCCCAG 5-10-5 90.0 295 405929 1337 1356 AAAGTTGGTCCCCAAAGTCC 5-10-5 85.5 296 399895 1340 1359 GCCAAAGTTGGTCCCCAAAG 3-14-3 84.6 36 395173 1340 1359 GCCAAAGTTGGTCCCCAAAG 5-10-5 94.8 36 405930 1342 1361 CGGCCAAAGTTGGTCCCCAA 5-10-5 92.9 297 405931 1344 1363 AGCGGCCAAAGTTGGTCCCC 5-10-5 88.8 298 405932 1346 1365 ACAGCGGCCAAAGTTGGTCC 5-10-5 90.6 299 405933 1348 1367 ACACAGCGGCCAAAGTTGGT 5-10-5 NA 300 405934 1350 1369 CCACACAGCGGCCAAAGTTG 5-10-5 92.7 301 405935 1354 1373 AGGTCCACACAGCGGCCAAA 5-10-5 86.0 302 410736 1356 1375 AGAGGTCCACACAGCGGCCA 5-10-5 73.8 303 405936 1358 1377 AAAGAGGTCCACACAGCGGC 5-10-5 93.9 304 410768 1380 1399 CAATGATGTCCTCCCCTGGG 5-10-5 79.7 305 405937 1387 1406 GAGGCACCAATGATGTCCTC 5-10-5 77.7 306 405938 1391 1410 GCTGGAGGCACCAATGATGT 5-10-5 75.5 307 405939 1393 1412 TCGCTGGAGGCACCAATGAT 5-10-5 70.6 308 405940 1395 1414 AGTCGCTGGAGGCACCAATG 5-10-5 84.4 309 405941 1397 1416 GCAGTCGCTGGAGGCACCAA 5-10-5 90.1 310 399804 1400 1419 GCTGCAGTCGCTGGAGGCAC 5-10-5 80.8 40 399960 1400 1419 GCTGCAGTCGCTGGAGGCAC 3-14-3 82.4 40 405942 1402 1421 GTGCTGCAGTCGCTGGAGGC 5-10-5 69.8 311 405943 1404 1423 AGGTGCTGCAGTCGCTGGAG 5-10-5 83.6 312 410737 1406 1425 GCAGGTGCTGCAGTCGCTGG 5-10-5 49.8 313 405944 1407 1426 AGCAGGTGCTGCAGTCGCTG 5-10-5 80.7 314 405945 1409 1428 AAAGCAGGTGCTGCAGTCGC 5-10-5 41.1 315 405946 1413 1432 ACACAAAGCAGGTGCTGCAG 5-10-5 70.4 316 405947 1415 1434 TGACACAAAGCAGGTGCTGC 5-10-5 72.5 317 410769 1425 1444 TCCCACTCTGTGACACAAAG 5-10-5 84.9 318 405948 1467 1486 TCATGGCTGCAATGCCAGCC 5-10-5 45.9 319 399806 1470 1489 GCATCATGGCTGCAATGCCA 5-10-5 82.8 42 399962 1470 1489 GCATCATGGCTGCAATGCCA 3-14-3 86.1 42 405949 1472 1491 CAGCATCATGGCTGCAATGC 5-10-5 84.9 320 405950 1474 1493 GACAGCATCATGGCTGCAAT 5-10-5 76.9 321 405951 1476 1495 CAGACAGCATCATGGCTGCA 5-10-5 75.4 322 405952 1480 1499 TCGGCAGACAGCATCATGGC 5-10-5 85.1 323 410738 1482 1501 GCTCGGCAGACAGCATCATG 5-10-5 68.4 324 405953 1484 1503 CGGCTCGGCAGACAGCATCA 5-10-5 93.6 325 405954 1486 1505 TCCGGCTCGGCAGACAGCAT 5-10-5 87.5 326 410770 1500 1519 CGGCCAGGGTGAGCTCCGGC 5-10-5 68.5 327 405955 1513 1532 CTCTGCCTCAACTCGGCCAG 5-10-5 94.2 328 405956 1515 1534 GTCTCTGCCTCAACTCGGCC 5-10-5 94.3 329 405958 1519 1538 ATCAGTCTCTGCCTCAACTC 5-10-5 80.3 331 405959 1521 1540 GGATCAGTCTCTGCCTCAAC 5-10-5 95.4 332 405960 1523 1542 GTGGATCAGTCTCTGCCTCA 5-10-5 90.1 333 405961 1525 1544 AAGTGGATCAGTCTCTGCCT 5-10-5 88.8 334 405962 1528 1547 GAGAAGTGGATCAGTCTCTG 5-10-5 56.1 335 410739 1530 1549 CAGAGAAGTGGATCAGTCTC 5-10-5 NA 336 405963 1532 1551 GGCAGAGAAGTGGATCAGTC 5-10-5 59.7 337 405964 1536 1555 CTTTGGCAGAGAAGTGGATC 5-10-5 45.3 338 405965 1541 1560 GACATCTTTGGCAGAGAAGT 5-10-5 55.9 339 405966 1543 1562 ATGACATCTTTGGCAGAGAA 5-10-5 51.3 340 405967 1547 1566 ATTGATGACATCTTTGGCAG 5-10-5 64.4 341 405968 1549 1568 TCATTGATGACATCTTTGGC 5-10-5 74.6 342 399967 1552 1571 GCCTCATTGATGACATCTTT 3-14-3 80.1 48 405969 1554 1573 AGGCCTCATTGATGACATCT 5-10-5 63.0 343 405970 1556 1575 CCAGGCCTCATTGATGACAT 5-10-5 66.8 344 410740 1558 1577 AACCAGGCCTCATTGATGAC 5-10-5 46.8 345 405885 1560 1579 GGAACCAGGCCTCATTGATG 5-10-5 73.4 346 410596 1562 1581 AGGGAACCAGGCCTCATTGA 3-14-3 NA 348 410672 1562 1581 AGGGAACCAGGCCTCATTGA 2-13-5 NA 348 405887 1562 1581 AGGGAACCAGGCCTCATTGA 5-10-5 NA 348 410597 1563 1582 CAGGGAACCAGGCCTCATTG 3-14-3 NA 349 405888 1563 1582 CAGGGAACCAGGCCTCATTG 5-10-5 NA 349 410673 1563 1582 CAGGGAACCAGGCCTCATTG 2-13-5 NA 349 410674 1564 1583 TCAGGGAACCAGGCCTCATT 2-13-5 60.7 49 399812 1564 1583 TCAGGGAACCAGGCCTCATT 5-10-5 68.7 49 399968 1564 1583 TCAGGGAACCAGGCCTCATT 3-14-3 84.5 49 410598 1565 1584 CTCAGGGAACCAGGCCTCAT 3-14-3 74.5 350 410675 1565 1584 CTCAGGGAACCAGGCCTCAT 2-13-5 76.9 350 405889 1565 1584 CTCAGGGAACCAGGCCTCAT 5-10-5 80.2 350 410599 1566 1585 CCTCAGGGAACCAGGCCTCA 3-14-3 70.6 351 410676 1566 1585 CCTCAGGGAACCAGGCCTCA 2-13-5 77.8 351 405890 1566 1585 CCTCAGGGAACCAGGCCTCA 5-10-5 87.7 351 410600 1567 1586 TCCTCAGGGAACCAGGCCTC 3-14-3 76.5 352 410677 1567 1586 TCCTCAGGGAACCAGGCCTC 2-13-5 88.4 352 405891 1567 1586 TCCTCAGGGAACCAGGCCTC 5-10-5 96.1 352 405892 1568 1587 GTCCTCAGGGAACCAGGCCT 5-10-5 71.4 353 410601 1568 1587 GTCCTCAGGGAACCAGGCCT 3-14-3 72.7 353 410678 1568 1587 GTCCTCAGGGAACCAGGCCT 2-13-5 75.0 353 395178 1569 1588 GGTCCTCAGGGAACCAGGCC 5-10-5 76.7 50 410679 1569 1588 GGTCCTCAGGGAACCAGGCC 2-13-5 77.6 50 399900 1569 1588 GGTCCTCAGGGAACCAGGCC 3-14-3 92.0 50 408653 1570 1589 TGGTCCTCAGGGAACCAGGC 5-10-5 42.5 354 410602 1570 1589 TGGTCCTCAGGGAACCAGGC 3-14-3 66.5 354 410680 1570 1589 TGGTCCTCAGGGAACCAGGC 2-13-5 71.6 354 410603 1571 1590 CTGGTCCTCAGGGAACCAGG 3-14-3 40.8 355 410681 1571 1590 CTGGTCCTCAGGGAACCAGG 2-13-5 44.4 355 405971 1571 1590 CTGGTCCTCAGGGAACCAGG 5-10-5 65.5 355 410540 1572 1591 GCTGGTCCTCAGGGAACCAG 5-10-5 50.9 356 410682 1572 1591 GCTGGTCCTCAGGGAACCAG 2-13-5 54.1 356 410604 1572 1591 GCTGGTCCTCAGGGAACCAG 3-14-3 62.5 356 405972 1573 1592 CGCTGGTCCTCAGGGAACCA 5-10-5 77.6 357 405973 1578 1597 GTACCCGCTGGTCCTCAGGG 5-10-5 83.8 358 405974 1580 1599 CAGTACCCGCTGGTCCTCAG 5-10-5 89.0 359 399813 1583 1602 GGTCAGTACCCGCTGGTCCT 5-10-5 72.4 51 399969 1583 1602 GGTCAGTACCCGCTGGTCCT 3-14-3 93.4 51 410771 1628 1647 ACCTGCCCCATGGGTGCTGG 5-10-5 NA 360 395181 1640 1659 AAACAGCTGCCAACCTGCCC 5-10-5 87.5 52 405975 1642 1661 CAAAACAGCTGCCAACCTGC 5-10-5 77.4 361 405976 1647 1666 TCCTGCAAAACAGCTGCCAA 5-10-5 81.2 362 405977 1649 1668 AGTCCTGCAAAACAGCTGCC 5-10-5 89.5 363 410772 1660 1679 GCTGACCATACAGTCCTGCA 5-10-5 91.2 364 405978 1672 1691 GGCCCCGAGTGTGCTGACCA 5-10-5 95.3 365 399904 1675 1694 GTAGGCCCCGAGTGTGCTGA 3-14-3 92.6 54 405979 1677 1696 GTGTAGGCCCCGAGTGTGCT 5-10-5 85.5 366 405980 1679 1698 CCGTGTAGGCCCCGAGTGTG 5-10-5 86.5 367 405981 1681 1700 ATCCGTGTAGGCCCCGAGTG 5-10-5 77.8 368 410741 1683 1702 CCATCCGTGTAGGCCCCGAG 5-10-5 78.4 369 405982 1685 1704 GGCCATCCGTGTAGGCCCCG 5-10-5 86.7 370 405983 1687 1706 GTGGCCATCCGTGTAGGCCC 5-10-5 73.1 371 405984 1735 1754 CTGGAGCAGCTCAGCAGCTC 5-10-5 83.4 372 405985 1737 1756 AACTGGAGCAGCTCAGCAGC 5-10-5 50.7 373 405986 1742 1761 GGAGAAACTGGAGCAGCTCA 5-10-5 75.6 374 405987 1744 1763 CTGGAGAAACTGGAGCAGCT 5-10-5 88.0 375 399905 1812 1831 CGTTGTGGGCCCGGCAGACC 3-14-3 91.2 57 395183 1812 1831 CGTTGTGGGCCCGGCAGACC 5-10-5 93.1 57 410541 1849 1868 CACCTGGCAATGGCGTAGAC 5-10-5 46.1 376 410605 1849 1868 CACCTGGCAATGGCGTAGAC 3-14-3 67.4 376 410683 1849 1868 CACCTGGCAATGGCGTAGAC 2-13-5 71.8 376 410606 1850 1869 GCACCTGGCAATGGCGTAGA 3-14-3 74.3 377 410542 1850 1869 GCACCTGGCAATGGCGTAGA 5-10-5 75.5 377 410684 1850 1869 GCACCTGGCAATGGCGTAGA 2-13-5 78.7 377 410543 1851 1870 AGCACCTGGCAATGGCGTAG 5-10-5 76.4 378 410685 1851 1870 AGCACCTGGCAATGGCGTAG 2-13-5 76.9 378 410607 1851 1870 AGCACCTGGCAATGGCGTAG 3-14-3 77.1 378 410544 1852 1871 CAGCACCTGGCAATGGCGTA 5-10-5 62.7 379 410608 1852 1871 CAGCACCTGGCAATGGCGTA 3-14-3 69.6 379 410686 1852 1871 CAGCACCTGGCAATGGCGTA 2-13-5 81.0 379 410545 1853 1872 GCAGCACCTGGCAATGGCGT 5-10-5 75.5 380 410687 1853 1872 GCAGCACCTGGCAATGGCGT 2-13-5 79.2 380 410609 1853 1872 GCAGCACCTGGCAATGGCGT 3-14-3 83.2 380 410610 1854 1873 GGCAGCACCTGGCAATGGCG 3-14-3 67.5 381 410688 1854 1873 GGCAGCACCTGGCAATGGCG 2-13-5 89.3 381 405988 1854 1873 GGCAGCACCTGGCAATGGCG 5-10-5 95.9 381 410546 1855 1874 AGGCAGCACCTGGCAATGGC 5-10-5 74.8 382 410689 1855 1874 AGGCAGCACCTGGCAATGGC 2-13-5 83.9 382 410611 1855 1874 AGGCAGCACCTGGCAATGGC 3-14-3 88.2 382 410612 1856 1875 CAGGCAGCACCTGGCAATGG 3-14-3 72.8 383 410690 1856 1875 CAGGCAGCACCTGGCAATGG 2-13-5 74.8 383 405989 1856 1875 CAGGCAGCACCTGGCAATGG 5-10-5 88.1 383 410547 1857 1876 GCAGGCAGCACCTGGCAATG 5-10-5 63.0 384 410691 1857 1876 GCAGGCAGCACCTGGCAATG 2-13-5 70.2 384 410613 1857 1876 GCAGGCAGCACCTGGCAATG 3-14-3 76.7 384 395184 1858 1877 AGCAGGCAGCACCTGGCAAT 5-10-5 68.5 58 410692 1858 1877 AGCAGGCAGCACCTGGCAAT 2-13-5 69.1 58 399906 1858 1877 AGCAGGCAGCACCTGGCAAT 3-14-3 94.2 58 410614 1859 1878 TAGCAGGCAGCACCTGGCAA 3-14-3 36.6 385 410548 1859 1878 TAGCAGGCAGCACCTGGCAA 5-10-5 75.0 385 410693 1859 1878 TAGCAGGCAGCACCTGGCAA 2-13-5 82.0 385 405990 1860 1879 GTAGCAGGCAGCACCTGGCA 5-10-5 96.8 386 410773 1905 1924 TGGCCTCAGCTGGTGGAGCT 5-10-5 61.3 387 410549 1915 1934 GTCCCCATGCTGGCCTCAGC 5-10-5 63.0 388 410615 1915 1934 GTCCCCATGCTGGCCTCAGC 3-14-3 72.5 388 410694 1915 1934 GTCCCCATGCTGGCCTCAGC 2-13-5 74.8 388 410550 1916 1935 GGTCCCCATGCTGGCCTCAG 5-10-5 69.4 389 410616 1916 1935 GGTCCCCATGCTGGCCTCAG 3-14-3 80.7 389 410695 1916 1935 GGTCCCCATGCTGGCCTCAG 2-13-5 85.5 389 410551 1917 1936 GGGTCCCCATGCTGGCCTCA 5-10-5 68.7 390 410696 1917 1936 GGGTCCCCATGCTGGCCTCA 2-13-5 86.3 390 410617 1917 1936 GGGTCCCCATGCTGGCCTCA 3-14-3 86.5 390 410552 1918 1937 CGGGTCCCCATGCTGGCCTC 5-10-5 77.9 391 410618 1918 1937 CGGGTCCCCATGCTGGCCTC 3-14-3 86.7 391 410697 1918 1937 CGGGTCCCCATGCTGGCCTC 2-13-5 87.0 391 410553 1919 1938 ACGGGTCCCCATGCTGGCCT 5-10-5 72.8 392 410619 1919 1938 ACGGGTCCCCATGCTGGCCT 3-14-3 84.1 392 410698 1919 1938 ACGGGTCCCCATGCTGGCCT 2-13-5 87.0 392 410699 1920 1939 CACGGGTCCCCATGCTGGCC 2-13-5 79.5 59 395185 1920 1939 CACGGGTCCCCATGCTGGCC 5-10-5 92.7 59 399907 1920 1939 CACGGGTCCCCATGCTGGCC 3-14-3 93.7 59 410620 1921 1940 ACACGGGTCCCCATGCTGGC 3-14-3 74.4 393 410554 1921 1940 ACACGGGTCCCCATGCTGGC 5-10-5 79.7 393 410700 1921 1940 ACACGGGTCCCCATGCTGGC 2-13-5 84.4 393 410621 1922 1941 GACACGGGTCCCCATGCTGG 3-14-3 76.0 394 410701 1922 1941 GACACGGGTCCCCATGCTGG 2-13-5 83.4 394 405991 1922 1941 GACACGGGTCCCCATGCTGG 5-10-5 91.9 394 410555 1923 1942 GGACACGGGTCCCCATGCTG 5-10-5 77.0 395 410622 1923 1942 GGACACGGGTCCCCATGCTG 3-14-3 79.8 395 410702 1923 1942 GGACACGGGTCCCCATGCTG 2-13-5 85.0 395 410703 1924 1943 TGGACACGGGTCCCCATGCT 2-13-5 70.4 396 410623 1924 1943 TGGACACGGGTCCCCATGCT 3-14-3 78.9 396 405992 1924 1943 TGGACACGGGTCCCCATGCT 5-10-5 89.3 396 410704 1925 1944 GTGGACACGGGTCCCCATGC 2-13-5 78.6 397 410556 1925 1944 GTGGACACGGGTCCCCATGC 5-10-5 81.4 397 410624 1925 1944 GTGGACACGGGTCCCCATGC 3-14-3 82.8 397 410625 1926 1945 AGTGGACACGGGTCCCCATG 3-14-3 64.4 398 410705 1926 1945 AGTGGACACGGGTCCCCATG 2-13-5 74.7 398 405993 1926 1945 AGTGGACACGGGTCCCCATG 5-10-5 85.5 398 410706 1927 1946 CAGTGGACACGGGTCCCCAT 2-13-5 70.8 399 410557 1927 1946 CAGTGGACACGGGTCCCCAT 5-10-5 74.2 399 410626 1927 1946 CAGTGGACACGGGTCCCCAT 3-14-3 76.8 399 410627 1928 1947 GCAGTGGACACGGGTCCCCA 3-14-3 71.4 400 410707 1928 1947 GCAGTGGACACGGGTCCCCA 2-13-5 72.8 400 405994 1928 1947 GCAGTGGACACGGGTCCCCA 5-10-5 95.2 400 410708 1929 1948 GGCAGTGGACACGGGTCCCC 2-13-5 79.0 401 410628 1929 1948 GGCAGTGGACACGGGTCCCC 3-14-3 88.1 401 410558 1929 1948 GGCAGTGGACACGGGTCCCC 5-10-5 88.9 401 410629 1930 1949 TGGCAGTGGACACGGGTCCC 3-14-3 66.5 402 410709 1930 1949 TGGCAGTGGACACGGGTCCC 2-13-5 66.9 402 405995 1930 1949 TGGCAGTGGACACGGGTCCC 5-10-5 92.4 402 410630 1931 1950 GTGGCAGTGGACACGGGTCC 3-14-3 54.1 403 410710 1931 1950 GTGGCAGTGGACACGGGTCC 2-13-5 61.4 403 410559 1931 1950 GTGGCAGTGGACACGGGTCC 5-10-5 77.5 403 410711 1932 1951 GGTGGCAGTGGACACGGGTC 2-13-5 NA 404 410631 1932 1951 GGTGGCAGTGGACACGGGTC 3-14-3 NA 404 410560 1932 1951 GGTGGCAGTGGACACGGGTC 5-10-5 42.5 404 410712 1933 1952 TGGTGGCAGTGGACACGGGT 2-13-5 NA 405 410632 1933 1952 TGGTGGCAGTGGACACGGGT 3-14-3 NA 405 410561 1933 1952 TGGTGGCAGTGGACACGGGT 5-10-5 NA 405 410774 1936 1955 TGTTGGTGGCAGTGGACACG 5-10-5 47.8 406 410775 1962 1981 AGCTGCAGCCTGTGAGGACG 5-10-5 36.3 407 410776 1990 2009 GTGCCAAGGTCCTCCACCTC 5-10-5 77.3 408 410777 2010 2029 TCAGCACAGGCGGCTTGTGG 5-10-5 40.0 409 410778 2040 2059 CCACGCACTGGTTGGGCTGA 5-10-5 52.7 410 399908 2100 2119 CTTTGCATTCCAGACCTGGG 3-14-3 74.9 60 410713 2100 2119 CTTTGCATTCCAGACCTGGG 2-13-5 84.9 60 395186 2100 2119 CTTTGCATTCCAGACCTGGG 5-10-5 93.7 60 410714 2101 2120 ACTTTGCATTCCAGACCTGG 2-13-5 82.4 411 410633 2101 2120 ACTTTGCATTCCAGACCTGG 3-14-3 83.8 411 410562 2101 2120 ACTTTGCATTCCAGACCTGG 5-10-5 87.3 411 410715 2102 2121 GACTTTGCATTCCAGACCTG 2-13-5 80.5 412 410634 2102 2121 GACTTTGCATTCCAGACCTG 3-14-3 81.8 412 405996 2102 2121 GACTTTGCATTCCAGACCTG 5-10-5 91.0 412 410563 2103 2122 TGACTTTGCATTCCAGACCT 5-10-5 75.4 413 410635 2103 2122 TGACTTTGCATTCCAGACCT 3-14-3 75.9 413 410716 2103 2122 TGACTTTGCATTCCAGACCT 2-13-5 88.3 413 410636 2104 2123 TTGACTTTGCATTCCAGACC 3-14-3 61.3 414 410564 2104 2123 TTGACTTTGCATTCCAGACC 5-10-5 71.3 414 410717 2104 2123 TTGACTTTGCATTCCAGACC 2-13-5 76.7 414 410718 2105 2124 CTTGACTTTGCATTCCAGAC 2-13-5 55.8 61 399973 2105 2124 CTTGACTTTGCATTCCAGAC 3-14-3 82.1 61 399817 2105 2124 CTTGACTTTGCATTCCAGAC 5-10-5 93.6 61 405997 2107 2126 TCCTTGACTTTGCATTCCAG 5-10-5 95.6 415 410779 2120 2139 CGGGATTCCATGCTCCTTGA 5-10-5 79.7 416 410780 2150 2169 GCAGGCCACGGTCACCTGCT 5-10-5 60.9 417 410781 2187 2206 GGAGGGCACTGCAGCCAGTC 5-10-5 66.0 418 410719 2305 2324 CAGATGGCAACGGCTGTCAC 2-13-5 61.0 419 410637 2305 2324 CAGATGGCAACGGCTGTCAC 3-14-3 66.0 419 410565 2305 2324 CAGATGGCAACGGCTGTCAC 5-10-5 71.6 419 410638 2306 2325 GCAGATGGCAACGGCTGTCA 3-14-3 71.9 420 410720 2306 2325 GCAGATGGCAACGGCTGTCA 2-13-5 75.1 420 410566 2306 2325 GCAGATGGCAACGGCTGTCA 5-10-5 77.7 420 410639 2307 2326 AGCAGATGGCAACGGCTGTC 3-14-3 67.7 421 410721 2307 2326 AGCAGATGGCAACGGCTGTC 2-13-5 68.8 421 410567 2307 2326 AGCAGATGGCAACGGCTGTC 5-10-5 72.1 421 410568 2308 2327 CAGCAGATGGCAACGGCTGT 5-10-5 54.2 422 410640 2308 2327 CAGCAGATGGCAACGGCTGT 3-14-3 57.7 422 410722 2308 2327 CAGCAGATGGCAACGGCTGT 2-13-5 59.2 422 410569 2309 2328 GCAGCAGATGGCAACGGCTG 5-10-5 57.7 423 410641 2309 2328 GCAGCAGATGGCAACGGCTG 3-14-3 66.1 423 410723 2309 2328 GCAGCAGATGGCAACGGCTG 2-13-5 81.1 423 399909 2310 2329 GGCAGCAGATGGCAACGGCT 3-14-3 69.9 62 410724 2310 2329 GGCAGCAGATGGCAACGGCT 2-13-5 87.4 62 395187 2310 2329 GGCAGCAGATGGCAACGGCT 5-10-5 92.7 62 410642 2311 2330 CGGCAGCAGATGGCAACGGC 3-14-3 70.4 424 410725 2311 2330 CGGCAGCAGATGGCAACGGC 2-13-5 75.6 424 410570 2311 2330 CGGCAGCAGATGGCAACGGC 5-10-5 78.3 424 410571 2312 2331 CCGGCAGCAGATGGCAACGG 5-10-5 52.8 425 410643 2312 2331 CCGGCAGCAGATGGCAACGG 3-14-3 59.8 425 410726 2312 2331 CCGGCAGCAGATGGCAACGG 2-13-5 69.8 425 410644 2313 2332 TCCGGCAGCAGATGGCAACG 3-14-3 59.0 426 410727 2313 2332 TCCGGCAGCAGATGGCAACG 2-13-5 68.3 426 405998 2313 2332 TCCGGCAGCAGATGGCAACG 5-10-5 75.6 426 410572 2314 2333 CTCCGGCAGCAGATGGCAAC 5-10-5 40.9 427 410728 2314 2333 CTCCGGCAGCAGATGGCAAC 2-13-5 56.4 427 410645 2314 2333 CTCCGGCAGCAGATGGCAAC 3-14-3 70.2 427 410729 2315 2334 GCTCCGGCAGCAGATGGCAA 2-13-5 67.7 428 410573 2315 2334 GCTCCGGCAGCAGATGGCAA 5-10-5 71.8 428 410646 2315 2334 GCTCCGGCAGCAGATGGCAA 3-14-3 72.8 428 410782 2325 2344 CCAGGTGCCGGCTCCGGCAG 5-10-5 43.2 429 410783 2335 2354 GAGGCCTGCGCCAGGTGCCG 5-10-5 63.7 430 399819 2509 2528 CCCACTCAAGGGCCAGGCCA 5-10-5 93.4 65 399912 2597 2616 ATGCCCCACAGTGAGGGAGG 3-14-3 84.3 66 405893 2828 2847 ATGAGGGCCATCAGCACCTT 5-10-5 93.9 431 405894 2829 2848 GATGAGGGCCATCAGCACCT 5-10-5 93.9 432 405895 2830 2849 AGATGAGGGCCATCAGCACC 5-10-5 91.8 433 405896 2831 2850 GAGATGAGGGCCATCAGCAC 5-10-5 88.5 434 395194 2832 2851 GGAGATGAGGGCCATCAGCA 5-10-5 90.0 69 399916 2832 2851 GGAGATGAGGGCCATCAGCA 3-14-3 94.5 69 405897 2833 2852 TGGAGATGAGGGCCATCAGC 5-10-5 85.1 435 405898 2834 2853 CTGGAGATGAGGGCCATCAG 5-10-5 86.9 436 405899 2835 2854 GCTGGAGATGAGGGCCATCA 5-10-5 91.5 437 405900 2836 2855 AGCTGGAGATGAGGGCCATC 5-10-5 83.7 438 405901 2902 2921 GCTAGATGCCATCCAGAAAG 5-10-5 88.4 439 405902 2903 2922 GGCTAGATGCCATCCAGAAA 5-10-5 89.3 440 405903 2904 2923 TGGCTAGATGCCATCCAGAA 5-10-5 92.6 441 405904 2905 2924 CTGGCTAGATGCCATCCAGA 5-10-5 90.4 442 399821 2906 2925 TCTGGCTAGATGCCATCCAG 5-10-5 91.7 71 405905 2907 2926 CTCTGGCTAGATGCCATCCA 5-10-5 90.7 443 405906 2908 2927 CCTCTGGCTAGATGCCATCC 5-10-5 93.3 444 405907 2909 2928 GCCTCTGGCTAGATGCCATC 5-10-5 90.2 445 405908 2910 2929 AGCCTCTGGCTAGATGCCAT 5-10-5 83.3 446 399978 2988 3007 AGCCTGGCATAGAGCAGAGT 3-14-3 96.4 73

Antisense oligonucleotides that exhibited less than 30% inhibition of PCSK9 mRNA levels were marked with “NA”.

Antisense oligonucleotides with the following ISIS Nos exhibited at least 80% inhibition of PCSK9 mRNA levels: 395152, 395153, 395155, 395158, 395163, 395165, 395166, 395168, 395173, 395181, 395183, 395185, 395186, 395187, 395194, 399798, 399801, 399804, 399806, 399817, 399819, 399821, 399877, 399879, 399887, 399888, 399891, 399895, 399900, 399904, 399905, 399906, 399907, 399912, 399916, 399954, 399960, 399962, 399967, 399968, 399969, 399973, 399978, 405861, 405862, 405863, 405864, 405865, 405866, 405869, 405870, 405871, 405872, 405873, 405874, 405875, 405876, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405889, 405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405900, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405908, 405909, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921, 405922, 405923, 405924, 405925, 405926, 405927, 405928, 405929, 405930, 405931, 405932, 405934, 405935, 405936, 405940, 405941, 405943, 405944, 405949, 405952, 405953, 405954, 405955, 405956, 405958, 405959, 405960, 405961, 405973, 405974, 405976, 405977, 405978, 405979, 405980, 405982, 405984, 405987, 405988, 405989, 405990, 405991, 405992, 405993, 405994, 405995, 405996, 405997, 406002, 406003, 406005, 406008, 406009, 406014, 406016, 406020, 406023, 406026, 406028, 406029, 406030, 406031, 406032, 406033, 406035, 406036, 406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410530, 410556, 410558, 410562, 410578, 410583, 410589, 410590, 410591, 410592, 410593, 410595, 410609, 410611, 410616, 410617, 410618, 410619, 410624, 410628, 410633, 410634, 410647, 410648, 410650, 410652, 410658, 410664, 410666, 410667, 410668, 410671, 410677, 410686, 410688, 410689, 410693, 410695, 410696, 410697, 410698, 410700, 410701, 410702, 410713, 410714, 410715, 410716, 410723, 410724, 410730, 410751, 410754, 410757, 410759, 410760, 410769, and 410772. The target segments to which these antisense oligonucleotides are targeted are active target segments. The target regions to which these antisense oligonucleotides are targeted are active target regions.

ISIS Nos 395152, 395155, 395158, 395165, 395166, 395168, 395173, 395181, 395183, 395185, 395186, 395187, 395194, 399801, 399817, 399819, 399821, 399877, 399879, 399887, 399888, 399891, 399900, 399904, 399905, 399906, 399907, 399916, 399954, 399962, 399969, 399978, 405861, 405862, 405864, 405869, 405871, 405872, 405873, 405874, 405875, 405876, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921, 405922, 405923, 405924, 405925, 405926, 405927, 405928, 405929, 405930, 405931, 405932, 405934, 405935, 405936, 405941, 405952, 405953, 405954, 405955, 405956, 405959, 405960, 405961, 405974, 405977, 405978, 405979, 405980, 405982, 405987, 405988, 405989, 405990, 405991, 405992, 405993, 405994, 405995, 405996, 405997, 406008, 406009, 406014, 406016, 406023, 406028, 406029, 406030, 406031, 406032, 406033, 406035, 406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410558, 410562, 410583, 410591, 410592, 410593, 410611, 410617, 410618, 410628, 410647, 410650, 410668, 410677, 410688, 410695, 410696, 410697, 410698, 410702, 410716, 410724, 410730, 410757, 410760, and 410772 each exhibited at least 85% inhibition of PCSK9 mRNA levels.

ISIS Nos 395152, 395155, 395158, 395165, 395166, 395168, 395173, 395183, 395185, 395186, 395187, 395194, 399801, 399817, 399819, 399821, 399877, 399879, 399887, 399888, 399891, 399900, 399904, 399905, 399906, 399907, 399916, 399954, 399969, 399978, 405861, 405862, 405864, 405873, 405874, 405875, 405876, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405891, 405893, 405894, 405895, 405899, 405903, 405904, 405905, 405906, 405907, 405910, 405913, 405914, 405916, 405923, 405924, 405925, 405926, 405927, 405928, 405930, 405932, 405934, 405936, 405941, 405953, 405955, 405956, 405959, 405960, 405978, 405988, 405990, 405991, 405994, 405995, 405996, 405997, 406008, 406009, 406014, 406023, 406028, 406029, 406030, 406033, 406035, 406039, 406040, 406041, 406042, 406043, 406044, 410592, 410647, 410668, 410760, and 410772 each exhibited at least 90% inhibition of PCSK9 mRNA levels.

ISIS Nos 395165, 395166, 399801, 399978, 405881, 405891, 405959, 405978, 405988, 405990, 405994, 405997, 406008, 406030, 406040, 406041, 406044, and 410760 each exhibited at least 95% inhibition of PCSK9 mRNA levels.

TABLE 18 Antisense inhibition of PCSK9 in human cells (Hep3B) targeting SEQ ID NO: 2 5′ Target 3′ Target Site to Site to SEQ SEQ ID SEQ ID % ID Isis No. NO: 2 NO: 2 Sequence 5′-3′ Motif inhibition NO: 410742 2433 2452 GCCTGGAGCTGACGGTGCCC 5-10-5 75.1 159 405999 2437 2456 GACCGCCTGGAGCTGACGGT 5-10-5 70.3 160 395151 2439 2458 AGGACCGCCTGGAGCTGACG 5-10-5 57.3 6 405861 2545 2564 AAGGCTAGCACCAGCTCCTC 5-10-5 90.5 162 405862 2546 2565 CAAGGCTAGCACCAGCTCCT 5-10-5 92.3 163 405863 2547 2566 GCAAGGCTAGCACCAGCTCC 5-10-5 82.6 164 405864 2548 2567 CGCAAGGCTAGCACCAGCTC 5-10-5 91.8 165 395152 2549 2568 ACGCAAGGCTAGCACCAGCT 5-10-5 93.3 7 405865 2550 2569 AACGCAAGGCTAGCACCAGC 5-10-5 84.6 166 405866 2551 2570 GAACGCAAGGCTAGCACCAG 5-10-5 81.8 167 405867 2552 2571 GGAACGCAAGGCTAGCACCA 5-10-5 74.0 168 405868 2553 2572 CGGAACGCAAGGCTAGCACC 5-10-5 76.8 169 399793 2556 2575 CCTCGGAACGCAAGGCTAGC 5-10-5 77.9 8 410743 2560 2579 TCCTCCTCGGAACGCAAGGC 5-10-5 76.7 170 410744 2585 2604 GTGCTCGGGTGCTTCGGCCA 5-10-5 76.9 171 410745 2605 2624 TGGAAGGTGGCTGTGGTTCC 5-10-5 41.7 172 395153 2619 2638 CCTTGGCGCAGCGGTGGAAG 5-10-5 81.1 9 410746 6444 6463 CGTAGGTGCCAGGCAACCTC 5-10-5 44.8 176 410747 6482 6501 TGACTGCGAGAGGTGGGTCT 5-10-5 NA 177 406003 6492 6511 CAGTGCGCTCTGACTGCGAG 5-10-5 84.2 178 406004 6494 6513 GGCAGTGCGCTCTGACTGCG 5-10-5 56.4 179 406005 6496 6515 CGGGCAGTGCGCTCTGACTG 5-10-5 83.4 180 406006 6499 6518 CGGCGGGCAGTGCGCTCTGA 5-10-5 71.0 181 410748 6528 6547 ATCCCCGGCGGGCAGCCTGG 5-10-5 56.0 182 406007 6532 6551 AGGTATCCCCGGCGGGCAGC 5-10-5 76.3 183 410574 6534 6553 TGAGGTATCCCCGGCGGGCA 3-14-3 69.1 184 410529 6534 6553 TGAGGTATCCCCGGCGGGCA 5-10-5 77.4 184 410647 6534 6553 TGAGGTATCCCCGGCGGGCA 2-13-5 90.7 184 410575 6535 6554 GTGAGGTATCCCCGGCGGGC 3-14-3 71.5 185 410648 6535 6554 GTGAGGTATCCCCGGCGGGC 2-13-5 82.8 185 410530 6535 6554 GTGAGGTATCCCCGGCGGGC 5-10-5 83.2 185 410649 6536 6555 GGTGAGGTATCCCCGGCGGG 2-13-5 53.5 186 410576 6536 6555 GGTGAGGTATCCCCGGCGGG 3-14-3 56.2 186 410531 6536 6555 GGTGAGGTATCCCCGGCGGG 5-10-5 63.6 186 410650 6537 6556 TGGTGAGGTATCCCCGGCGG 2-13-5 86.1 11 399877 6537 6556 TGGTGAGGTATCCCCGGCGG 3-14-3 92.6 11 395155 6537 6556 TGGTGAGGTATCCCCGGCGG 5-10-5 92.7 11 410577 6538 6557 TTGGTGAGGTATCCCCGGCG 3-14-3 62.4 187 410532 6538 6557 TTGGTGAGGTATCCCCGGCG 5-10-5 63.5 187 410651 6538 6557 TTGGTGAGGTATCCCCGGCG 2-13-5 75.5 187 410652 6539 6558 CTTGGTGAGGTATCCCCGGC 2-13-5 81.2 188 410578 6539 6558 CTTGGTGAGGTATCCCCGGC 3-14-3 83.3 188 406008 6539 6558 CTTGGTGAGGTATCCCCGGC 5-10-5 95.2 188 410653 6540 6559 TCTTGGTGAGGTATCCCCGG 2-13-5 68.9 189 410579 6540 6559 TCTTGGTGAGGTATCCCCGG 3-14-3 74.0 189 410533 6540 6559 TCTTGGTGAGGTATCCCCGG 5-10-5 79.0 189 410580 6541 6560 ATCTTGGTGAGGTATCCCCG 3-14-3 62.3 190 410654 6541 6560 ATCTTGGTGAGGTATCCCCG 2-13-5 65.2 190 406009 6541 6560 ATCTTGGTGAGGTATCCCCG 5-10-5 90.7 190 410534 6542 6561 GATCTTGGTGAGGTATCCCC 5-10-5 59.8 191 410581 6542 6561 GATCTTGGTGAGGTATCCCC 3-14-3 72.9 191 410655 6542 6561 GATCTTGGTGAGGTATCCCC 2-13-5 77.4 191 399794 6543 6562 GGATCTTGGTGAGGTATCCC 5-10-5 56.5 12 410656 6543 6562 GGATCTTGGTGAGGTATCCC 2-13-5 68.0 12 399950 6543 6562 GGATCTTGGTGAGGTATCCC 3-14-3 74.4 12 410535 6544 6563 AGGATCTTGGTGAGGTATCC 5-10-5 65.7 192 410582 6544 6563 AGGATCTTGGTGAGGTATCC 3-14-3 69.5 192 410657 6544 6563 AGGATCTTGGTGAGGTATCC 2-13-5 73.1 192 406010 6546 6565 GCAGGATCTTGGTGAGGTAT 5-10-5 56.9 193 406011 6548 6567 ATGCAGGATCTTGGTGAGGT 5-10-5 70.7 194 406012 6550 6569 ACATGCAGGATCTTGGTGAG 5-10-5 69.1 195 406013 6554 6573 GAAGACATGCAGGATCTTGG 5-10-5 77.4 196 395156 6557 6576 ATGGAAGACATGCAGGATCT 5-10-5 76.7 14 410749 6565 6584 AGAAGGCCATGGAAGACATG 5-10-5 59.4 197 410750 6575 6594 GAAGCCAGGAAGAAGGCCAT 5-10-5 57.5 198 399879 6583 6602 TTCACCAGGAAGCCAGGAAG 3-14-3 91.9 15 406014 6585 6604 TCTTCACCAGGAAGCCAGGA 5-10-5 94.0 199 406015 6590 6609 ACTCATCTTCACCAGGAAGC 5-10-5 78.8 200 406016 6592 6611 CCACTCATCTTCACCAGGAA 5-10-5 87.2 201 410730 6594 6613 CGCCACTCATCTTCACCAGG 5-10-5 85.4 202 406017 6596 6615 GTCGCCACTCATCTTCACCA 5-10-5 76.3 203 406018 6598 6617 AGGTCGCCACTCATCTTCAC 5-10-5 63.2 204 406019 6600 6619 GCAGGTCGCCACTCATCTTC 5-10-5 68.1 205 406020 6602 6621 CAGCAGGTCGCCACTCATCT 5-10-5 83.7 206 410731 6604 6623 TCCAGCAGGTCGCCACTCAT 5-10-5 72.9 207 395158 9130 9149 CCTCGATGTAGTCGACATGG 5-10-5 93.9 17 410752 9149 9168 GCAAAGACAGAGGAGTCCTC 5-10-5 76.3 209 406021 9207 9226 TTCATCCGCCCGGTACCGTG 5-10-5 79.6 210 406022 9209 9228 TATTCATCCGCCCGGTACCG 5-10-5 78.7 211 406023 9212 9231 TGGTATTCATCCGCCCGGTA 5-10-5 92.5 212 406024 9214 9233 GCTGGTATTCATCCGCCCGG 5-10-5 69.7 213 410732 9216 9235 GGGCTGGTATTCATCCGCCC 5-10-5 30.4 214 399935 14601 14620 AGGACCCAAGTCATCCTGCT 3-14-3 81.1 96 399936 14631 14650 GGCCATCAGCTGGCAATGCT 3-14-3 89.0 124 410753 14877 14896 ATACACCTCCACCAGGCTGC 5-10-5 72.8 315 406025 14888 14907 GTGTCTAGGAGATACACCTC 5-10-5 74.2 216 406026 14893 14912 TGCTGGTGTCTAGGAGATAC 5-10-5 80.5 217 405869 14918 14937 TCGATTTCCCGGTGGTCACT 5-10-5 87.6 218 405870 14919 14938 CTCGATTTCCCGGTGGTCAC 5-10-5 83.3 219 405871 14920 14939 CCTCGATTTCCCGGTGGTCA 5-10-5 87.0 220 405872 14921 14940 CCCTCGATTTCCCGGTGGTC 5-10-5 88.2 221 399798 14922 14941 GCCCTCGATTTCCCGGTGGT 5-10-5 84.3 22 399954 14922 14941 GCCCTCGATTTCCCGGTGGT 3-14-3 90.3 22 405873 14923 14942 TGCCCTCGATTTCCCGGTGG 5-10-5 90.0 222 405874 14924 14943 CTGCCCTCGATTTCCCGGTG 5-10-5 91.4 223 405875 14925 14944 CCTGCCCTCGATTTCCCGGT 5-10-5 93.5 224 405876 14926 14945 CCCTGCCCTCGATTTCCCGG 5-10-5 90.1 225 406027 14930 14949 ATGACCCTGCCCTCGATTTC 5-10-5 73.9 226 406028 14932 14951 CCATGACCCTGCCCTCGATT 5-10-5 92.3 227 406029 14934 14953 GACCATGACCCTGCCCTCGA 5-10-5 91.4 228 406030 14938 14957 CGGTGACCATGACCCTGCCC 5-10-5 95.6 229 406031 14940 14959 GTCGGTGACCATGACCCTGC 5-10-5 89.6 230 410733 14942 14961 AAGTCGGTGACCATGACCCT 5-10-5 65.7 231 406032 14944 14963 CGAAGTCGGTGACCATGACC 5-10-5 88.5 232 410754 14954 14973 GGCACATTCTCGAAGTCGGT 5-10-5 80.1 233 395163 14979 14998 GTGGAAGCGGGTCCCGTCCT 5-10-5 83.9 25 410756 15254 15273 GGTGGGTGCCATGACTGTCA 5-10-5 67.1 235 406033 15261 15280 CCTGCCAGGTGGGTGCCATG 5-10-5 91.2 237 406034 15266 15285 CCACCCCTGCCAGGTGGGTG 5-10-5 59.3 238 406035 15271 15290 GCTGACCACCCCTGCCAGGT 5-10-5 91.7 239 410757 15279 15298 TCCCGGCCGCTGACCACCCC 5-10-5 88.9 240 406036 15283 15302 GGCATCCCGGCCGCTGACCA 5-10-5 84.0 241 406037 15286 15305 GCCGGCATCCCGGCCGCTGA 5-10-5 55.9 242 410536 15291 15310 GCCACGCCGGCATCCCGGCC 5-10-5 63.7 243 410658 15291 15310 GCCACGCCGGCATCCCGGCC 2-13-5 82.0 243 410583 15291 15310 GCCACGCCGGCATCCCGGCC 3-14-3 87.3 243 410537 15292 15311 GGCCACGCCGGCATCCCGGC 5-10-5 58.5 244 410659 15292 15311 GGCCACGCCGGCATCCCGGC 2-13-5 73.7 244 410584 15292 15311 GGCCACGCCGGCATCCCGGC 3-14-3 79.5 244 410660 15293 15312 TGGCCACGCCGGCATCCCGG 2-13-5 63.0 245 410585 15293 15312 TGGCCACGCCGGCATCCCGG 3-14-3 73.2 245 406038 15293 15312 TGGCCACGCCGGCATCCCGG 5-10-5 86.3 245 410661 15294 15313 TTGGCCACGCCGGCATCCCG 2-13-5 53.3 246 410586 15294 15313 TTGGCCACGCCGGCATCCCG 3-14-3 60.3 246 405877 15294 15313 TTGGCCACGCCGGCATCCCG 5-10-5 83.2 246 410587 15295 15314 CTTGGCCACGCCGGCATCCC 3-14-3 63.2 247 410662 15295 15314 CTTGGCCACGCCGGCATCCC 2-13-5 67.3 247 405878 15295 15314 CTTGGCCACGCCGGCATCCC 5-10-5 91.4 247 410588 15296 15315 CCTTGGCCACGCCGGCATCC 3-14-3 65.3 248 410663 15296 15315 CCTTGGCCACGCCGGCATCC 2-13-5 67.9 248 405879 15296 15315 CCTTGGCCACGCCGGCATCC 5-10-5 94.1 248 410589 15297 15316 CCCTTGGCCACGCCGGCATC 3-14-3 80.1 249 410664 15297 15316 CCCTTGGCCACGCCGGCATC 2-13-5 81.9 249 405880 15297 15316 CCCTTGGCCACGCCGGCATC 5-10-5 93.5 249 410665 15298 15317 ACCCTTGGCCACGCCGGCAT 2-13-5 78.7 28 399887 15298 15317 ACCCTTGGCCACGCCGGCAT 3-14-3 91.9 28 395165 15298 15317 ACCCTTGGCCACGCCGGCAT 5-10-5 96.7 28 410590 15299 15318 CACCCTTGGCCACGCCGGCA 3-14-3 82.2 250 410666 15299 15318 CACCCTTGGCCACGCCGGCA 2-13-5 82.7 250 405881 15299 15318 CACCCTTGGCCACGCCGGCA 5-10-5 98.3 250 410667 15300 15319 GCACCCTTGGCCACGCCGGC 2-13-5 84.7 251 410591 15300 15319 GCACCCTTGGCCACGCCGGC 3-14-3 86.3 251 405882 15300 15319 GCACCCTTGGCCACGCCGGC 5-10-5 91.6 251 410668 15301 15320 GGCACCCTTGGCCACGCCGG 2-13-5 91.4 252 405883 15301 15320 GGCACCCTTGGCCACGCCGG 5-10-5 92.5 252 410592 15301 15320 GGCACCCTTGGCCACGCCGG 3-14-3 93.4 252 410669 15302 15321 TGGCACCCTTGGCCACGCCG 2-13-5 69.0 253 410593 15302 15321 TGGCACCCTTGGCCACGCCG 3-14-3 88.8 253 405884 15302 15321 TGGCACCCTTGGCCACGCCG 5-10-5 90.8 253 410670 15303 15322 CTGGCACCCTTGGCCACGCC 2-13-5 74.9 254 410594 15303 15322 CTGGCACCCTTGGCCACGCC 3-14-3 75.2 254 410538 15303 15322 CTGGCACCCTTGGCCACGCC 5-10-5 78.2 254 410539 15304 15323 GCTGGCACCCTTGGCCACGC 5-10-5 61.9 255 410595 15304 15323 GCTGGCACCCTTGGCCACGC 3-14-3 83.1 255 410671 15304 15323 GCTGGCACCCTTGGCCACGC 2-13-5 83.3 255 410758 15309 15328 CGCATGCTGGCACCCTTGGC 5-10-5 73.6 256 406039 15330 15349 CAGTTGAGCACGCGCAGGCT 5-10-5 91.2 257 406040 15332 15351 GGCAGTTGAGCACGCGCAGG 5-10-5 96.5 258 399888 15334 15353 TTGGCAGTTGAGCACGCGCA 3-14-3 90.9 29 395166 15334 15353 TTGGCAGTTGAGCACGCGCA 5-10-5 96.2 29 406041 15336 15355 CCTTGGCAGTTGAGCACGCG 5-10-5 96.6 259 399801 15339 15358 TTCCCTTGGCAGTTGAGCAC 5-10-5 95.6 30 406042 15341 15360 CCTTCCCTTGGCAGTTGAGC 5-10-5 92.0 260 406043 15345 15364 GTGCCCTTCCCTTGGCAGTT 5-10-5 91.9 261 406044 15347 15366 CCGTGCCCTTCCCTTGGCAG 5-10-5 95.4 262 410759 15358 15377 GGTGCCGCTAACCGTGCCCT 5-10-5 83.7 263 406045 18591 18610 TGGCTTTTCCGAATAAACTC 5-10-5 88.4 266 395168 18593 18612 GCTGGCTTTTCCGAATAAAC 5-10-5 91.9 32 405909 18595 18614 CAGCTGGCTTTTCCGAATAA 5-10-5 80.3 267 405910 18597 18616 ACCAGCTGGCTTTTCCGAAT 5-10-5 90.9 268 405911 18599 18618 GGACCAGCTGGCTTTTCCGA 5-10-5 88.0 269 405912 18603 18622 GGCTGGACCAGCTGGCTTTT 5-10-5 78.2 270 410761 18614 18633 GTGGCCCCACAGGCTGGACC 5-10-5 53.7 271 410762 18627 18646 AGCAGCACCACCAGTGGCCC 5-10-5 57.6 272 410763 18649 18668 GCTGTACCCACCCGCCAGGG 5-10-5 68.8 273 410764 18695 18714 CGACCCCAGCCCTCGCCAGG 5-10-5 66.3 274 399891 18705 18724 GTGACCAGCACGACCCCAGC 3-14-3 91.1 33 405913 18707 18726 CGGTGACCAGCACGACCCCA 5-10-5 93.8 275 405914 18709 18728 AGCGGTGACCAGCACGACCC 5-10-5 90.4 276 405915 18711 18730 GCAGCGGTGACCAGCACGAC 5-10-5 86.8 277 405916 18713 18732 CGGCAGCGGTGACCAGCACG 5-10-5 91.3 278 410734 18714 18733 CCGGCAGCGGTGACCAGCAC 5-10-5 65.8 279 405917 18717 18736 TTGCCGGCAGCGGTGACCAG 5-10-5 62.0 280 405918 18719 18738 AGTTGCCGGCAGCGGTGACC 5-10-5 33.5 281 405919 18721 18740 GAAGTTGCCGGCAGCGGTGA 5-10-5 68.2 282 405920 18723 18742 CGGAAGTTGCCGGCAGCGGT 5-10-5 86.2 283 405921 18725 18744 CCCGGAAGTTGCCGGCAGCG 5-10-5 85.5 284 405922 18727 18746 GTCCCGGAAGTTGCCGGCAG 5-10-5 86.0 285 405923 19931 19950 GGCATTGGTGGCCCCAACTG 5-10-5 94.3 288 410767 19941 19960 GCTGGTCTTGGGCATTGGTG 5-10-5 65.1 289 405924 19954 19973 CCCAGGGTCACCGGCTGGTC 5-10-5 93.1 290 410735 19956 19975 TCCCCAGGGTCACCGGCTGG 5-10-5 78.1 291 405925 19958 19977 AGTCCCCAGGGTCACCGGCT 5-10-5 93.5 292 405926 19960 19979 AAAGTCCCCAGGGTCACCGG 5-10-5 94.6 293 405927 19964 19983 CCCCAAAGTCCCCAGGGTCA 5-10-5 94.5 294 405928 19969 19988 TTGGTCCCCAAAGTCCCCAG 5-10-5 90.0 295 405929 19973 19992 AAAGTTGGTCCCCAAAGTCC 5-10-5 85.5 296 399895 19976 19995 GCCAAAGTTGGTCCCCAAAG 3-14-3 84.6 36 395173 19976 19995 GCCAAAGTTGGTCCCCAAAG 5-10-5 94.8 36 405930 19978 19997 CGGCCAAAGTTGGTCCCCAA 5-10-5 92.9 297 405931 19980 19999 AGCGGCCAAAGTTGGTCCCC 5-10-5 88.8 298 405932 19982 20001 ACAGCGGCCAAAGTTGGTCC 5-10-5 90.6 299 405933 19984 20003 ACACAGCGGCCAAAGTTGGT 5-10-5 NA 300 405934 19986 20005 CCACACAGCGGCCAAAGTTG 5-10-5 92.7 301 405935 19990 20009 AGGTCCACACAGCGGCCAAA 5-10-5 86.0 302 410736 19992 20011 AGAGGTCCACACAGCGGCCA 5-10-5 73.8 303 405936 19994 20013 AAAGAGGTCCACACAGCGGC 5-10-5 93.9 304 410768 20016 20035 CAATGATGTCCTCCCCTGGG 5-10-5 79.7 305 405937 20023 20042 GAGGCACCAATGATGTCCTC 5-10-5 77.7 306 405938 20027 20046 GCTGGAGGCACCAATGATGT 5-10-5 75.5 307 405939 20029 20048 TCGCTGGAGGCACCAATGAT 5-10-5 70.6 308 405940 20031 20050 AGTCGCTGGAGGCACCAATG 5-10-5 84.4 309 405941 20033 20052 GCAGTCGCTGGAGGCACCAA 5-10-5 90.1 310 399804 20036 20055 GCTGCAGTCGCTGGAGGCAC 5-10-5 80.8 40 399960 20036 20055 GCTGCAGTCGCTGGAGGCAC 3-14-3 82.4 40 405942 20038 20057 GTGCTGCAGTCGCTGGAGGC 5-10-5 69.8 311 405943 20040 20059 AGGTGCTGCAGTCGCTGGAG 5-10-5 83.6 312 410737 20042 20061 GCAGGTGCTGCAGTCGCTGG 5-10-5 49.8 313 405944 20043 20062 AGCAGGTGCTGCAGTCGCTG 5-10-5 80.7 314 405945 20045 20064 AAAGCAGGTGCTGCAGTCGC 5-10-5 41.1 315 405946 20049 20068 ACACAAAGCAGGTGCTGCAG 5-10-5 70.4 316 405947 20051 20070 TGACACAAAGCAGGTGCTGC 5-10-5 72.5 317 410769 20061 20080 TCCCACTCTGTGACACAAAG 5-10-5 84.9 318 405949 20629 20648 CAGCATCATGGCTGCAATGC 5-10-5 84.9 320 405950 20631 20650 GACAGCATCATGGCTGCAAT 5-10-5 76.9 321 405951 20633 20652 CAGACAGCATCATGGCTGCA 5-10-5 75.4 322 405952 20637 20656 TCGGCAGACAGCATCATGGC 5-10-5 85.1 323 410738 20639 20658 GCTCGGCAGACAGCATCATG 5-10-5 68.4 324 405953 20641 20660 CGGCTCGGCAGACAGCATCA 5-10-5 93.6 325 405954 20643 20662 TCCGGCTCGGCAGACAGCAT 5-10-5 87.5 326 410770 20657 20676 CGGCCAGGGTGAGCTCCGGC 5-10-5 68.5 327 405955 20670 20689 CTCTGCCTCAACTCGGCCAG 5-10-5 94.2 328 405956 20672 20691 GTCTCTGCCTCAACTCGGCC 5-10-5 94.3 329 405958 20676 20695 ATCAGTCTCTGCCTCAACTC 5-10-5 80.3 331 405959 20678 20697 GGATCAGTCTCTGCCTCAAC 5-10-5 95.4 332 405960 20680 20699 GTGGATCAGTCTCTGCCTCA 5-10-5 90.1 333 405961 20682 20701 AAGTGGATCAGTCTCTGCCT 5-10-5 88.8 334 405962 20685 20704 GAGAAGTGGATCAGTCTCTG 5-10-5 56.1 335 410739 20687 20706 CAGAGAAGTGGATCAGTCTC 5-10-5 NA 336 405963 20689 20708 GGCAGAGAAGTGGATCAGTC 5-10-5 59.7 337 405964 20693 20712 CTTTGGCAGAGAAGTGGATC 5-10-5 45.3 338 405965 20698 20717 GACATCTTTGGCAGAGAAGT 5-10-5 55.9 339 405966 20700 20719 ATGACATCTTTGGCAGAGAA 5-10-5 51.3 340 405967 20704 20723 ATTGATGACATCTTTGGCAG 5-10-5 64.4 341 405968 20706 20725 TCATTGATGACATCTTTGGC 5-10-5 74.6 342 399967 20709 20728 GCCTCATTGATGACATCTTT 3-14-3 80.1 48 405969 20711 20730 AGGCCTCATTGATGACATCT 5-10-5 63.0 343 405970 20713 20732 CCAGGCCTCATTGATGACAT 5-10-5 66.8 344 410740 20715 20734 AACCAGGCCTCATTGATGAC 5-10-5 46.8 345 405885 20717 20736 GGAACCAGGCCTCATTGATG 5-10-5 73.4 346 410596 20719 20738 AGGGAACCAGGCCTCATTGA 3-14-3 NA 348 410672 20719 20738 AGGGAACCAGGCCTCATTGA 2-13-5 NA 348 405887 20719 20738 AGGGAACCAGGCCTCATTGA 5-10-5 NA 348 410597 20720 20739 CAGGGAACCAGGCCTCATTG 3-14-3 NA 349 405888 20720 20739 CAGGGAACCAGGCCTCATTG 5-10-5 NA 349 410673 20720 20739 CAGGGAACCAGGCCTCATTG 2-13-5 NA 349 410674 20721 20740 TCAGGGAACCAGGCCTCATT 2-13-5 60.7 49 399812 20721 20740 TCAGGGAACCAGGCCTCATT 5-10-5 68.7 49 399968 20721 20740 TCAGGGAACCAGGCCTCATT 3-14-3 84.5 49 410598 20722 20741 CTCAGGGAACCAGGCCTCAT 3-14-3 74.5 350 410675 20722 20741 CTCAGGGAACCAGGCCTCAT 2-13-5 76.9 350 405889 20722 20741 CTCAGGGAACCAGGCCTCAT 5-10-5 80.2 350 410599 20723 20742 CCTCAGGGAACCAGGCCTCA 3-14-3 70.6 351 410676 20723 20742 CCTCAGGGAACCAGGCCTCA 2-13-5 77.8 351 405890 20723 20742 CCTCAGGGAACCAGGCCTCA 5-10-5 87.7 351 410600 20724 20743 TCCTCAGGGAACCAGGCCTC 3-14-3 76.5 352 410677 20724 20743 TCCTCAGGGAACCAGGCCTC 2-13-5 88.4 352 405891 20724 20743 TCCTCAGGGAACCAGGCCTC 5-10-5 96.1 352 405892 20725 20744 GTCCTCAGGGAACCAGGCCT 5-10-5 71.4 353 410601 20725 20744 GTCCTCAGGGAACCAGGCCT 3-14-3 72.7 353 410678 20725 20744 GTCCTCAGGGAACCAGGCCT 2-13-5 75.0 353 395178 20726 20745 GGTCCTCAGGGAACCAGGCC 5-10-5 76.7 50 410679 20726 20745 GGTCCTCAGGGAACCAGGCC 2-13-5 77.6 50 399900 20726 20745 GGTCCTCAGGGAACCAGGCC 3-14-3 92.0 50 408653 20727 20746 TGGTCCTCAGGGAACCAGGC 5-10-5 42.5 354 410602 20727 20746 TGGTCCTCAGGGAACCAGGC 3-14-3 66.5 354 410680 20727 20746 TGGTCCTCAGGGAACCAGGC 2-13-5 71.6 354 410603 20728 20747 CTGGTCCTCAGGGAACCAGG 3-14-3 40.8 355 410681 20728 20747 CTGGTCCTCAGGGAACCAGG 2-13-5 44.4 355 405971 20728 20747 CTGGTCCTCAGGGAACCAGG 5-10-5 65.5 355 410540 20729 20748 GCTGGTCCTCAGGGAACCAG 5-10-5 50.9 356 410682 20729 20748 GCTGGTCCTCAGGGAACCAG 2-13-5 54.1 356 410604 20729 20748 GCTGGTCCTCAGGGAACCAG 3-14-3 62.5 356 405972 20730 20749 CGCTGGTCCTCAGGGAACCA 5-10-5 77.6 357 405973 20735 20754 GTACCCGCTGGTCCTCAGGG 5-10-5 83.8 358 405974 20737 20756 CAGTACCCGCTGGTCCTCAG 5-10-5 89.0 359 399813 20740 20759 GGTCAGTACCCGCTGGTCCT 5-10-5 72.4 51 399969 20740 20759 GGTCAGTACCCGCTGGTCCT 3-14-3 93.4 51 410771 20785 20804 ACCTGCCCCATGGGTGCTGG 5-10-5 NA 360 405975 21088 21107 CAAAACAGCTGCCAACCTGC 5-10-5 77.4 361 405976 21093 21112 TCCTGCAAAACAGCTGCCAA 5-10-5 81.2 362 405977 21095 21114 AGTCCTGCAAAACAGCTGCC 5-10-5 89.5 363 410772 21106 21125 GCTGACCATACAGTCCTGCA 5-10-5 91.2 264 405978 21118 21137 GGCCCCGAGTGTGCTGACCA 5-10-5 95.3 365 399904 21121 21140 GTAGGCCCCGAGTGTGCTGA 3-14-3 92.6 54 405979 21123 21142 GTGTAGGCCCCGAGTGTGCT 5-10-5 85.5 366 405980 21125 21144 CCGTGTAGGCCCCGAGTGTG 5-10-5 86.5 367 405981 21127 21146 ATCCGTGTAGGCCCCGAGTG 5-10-5 77.8 368 410741 21129 21148 CCATCCGTGTAGGCCCCGAG 5-10-5 78.4 369 405982 21131 21150 GGCCATCCGTGTAGGCCCCG 5-10-5 86.7 370 405983 21133 21152 GTGGCCATCCGTGTAGGCCC 5-10-5 73.1 371 405984 21181 21200 CTGGAGCAGCTCAGCAGCTC 5-10-5 83.4 372 405985 21183 21202 AACTGGAGCAGCTCAGCAGC 5-10-5 50.7 373 405986 21188 21207 GGAGAAACTGGAGCAGCTCA 5-10-5 75.6 374 405987 21190 21209 CTGGAGAAACTGGAGCAGCT 5-10-5 88.0 375 399868 21696 21715 GGCACTGCCCTTCCACCAAA 5-10-5 85.8 123 399905 22096 22115 CGTTGTGGGCCCGGCAGACC 3-14-3 91.2 57 395183 22096 22115 CGTTGTGGGCCCGGCAGACC 5-10-5 93.1 57 410541 22133 22152 CACCTGGCAATGGCGTAGAC 5-10-5 46.1 376 410605 22133 22152 CACCTGGCAATGGCGTAGAC 3-14-3 67.4 376 410683 22133 22152 CACCTGGCAATGGCGTAGAC 2-13-5 71.8 376 410606 22134 22153 GCACCTGGCAATGGCGTAGA 3-14-3 74.3 377 410542 22134 22153 GCACCTGGCAATGGCGTAGA 5-10-5 75.5 377 410684 22134 22153 GCACCTGGCAATGGCGTAGA 2-13-5 78.7 377 410543 22135 22154 AGCACCTGGCAATGGCGTAG 5-10-5 76.4 378 410685 22135 22154 AGCACCTGGCAATGGCGTAG 2-13-5 76.9 378 410607 22135 22154 AGCACCTGGCAATGGCGTAG 3-14-3 77.1 378 410544 22136 22155 CAGCACCTGGCAATGGCGTA 5-10-5 62.7 379 410608 22136 22155 CAGCACCTGGCAATGGCGTA 3-14-3 69.6 379 410686 22136 22155 CAGCACCTGGCAATGGCGTA 2-13-5 81.0 379 410545 22137 22156 GCAGCACCTGGCAATGGCGT 5-10-5 75.5 380 410687 22137 22156 GCAGCACCTGGCAATGGCGT 2-13-5 79.2 380 410609 22137 22156 GCAGCACCTGGCAATGGCGT 3-14-3 83.2 380 410610 22138 22157 GGCAGCACCTGGCAATGGCG 3-14-3 67.5 381 410688 22138 22157 GGCAGCACCTGGCAATGGCG 2-13-5 89.3 381 405988 22138 22157 GGCAGCACCTGGCAATGGCG 5-10-5 95.9 381 410546 22139 22158 AGGCAGCACCTGGCAATGGC 5-10-5 74.8 382 410689 22139 22158 AGGCAGCACCTGGCAATGGC 2-13-5 83.9 382 410611 22139 22158 AGGCAGCACCTGGCAATGGC 3-14-3 88.2 382 410612 22140 22159 CAGGCAGCACCTGGCAATGG 3-14-3 72.8 383 410690 22140 22159 CAGGCAGCACCTGGCAATGG 2-13-5 74.8 383 405989 22140 22159 CAGGCAGCACCTGGCAATGG 5-10-5 88.1 383 410547 22141 22160 GCAGGCAGCACCTGGCAATG 5-10-5 63.0 384 410691 22141 22160 GCAGGCAGCACCTGGCAATG 2-13-5 70.2 384 410613 22141 22160 GCAGGCAGCACCTGGCAATG 3-14-3 76.7 384 395184 22142 22161 AGCAGGCAGCACCTGGCAAT 5-10-5 68.5 58 410692 22142 22161 AGCAGGCAGCACCTGGCAAT 2-13-5 69.1 58 399906 22142 22161 AGCAGGCAGCACCTGGCAAT 3-14-3 94.2 58 410614 22143 22162 TAGCAGGCAGCACCTGGCAA 3-14-3 36.6 385 410548 22143 22162 TAGCAGGCAGCACCTGGCAA 5-10-5 75.0 385 410693 22143 22162 TAGCAGGCAGCACCTGGCAA 2-13-5 82.0 385 405990 22144 22163 GTAGCAGGCAGCACCTGGCA 5-10-5 96.8 386 410773 22189 22208 TGGCCTCAGCTGGTGGAGCT 5-10-5 61.3 387 410549 22199 22218 GTCCCCATGCTGGCCTCAGC 5-10-5 63.0 388 410615 22199 22218 GTCCCCATGCTGGCCTCAGC 3-14-3 72.5 388 410694 22199 22218 GTCCCCATGCTGGCCTCAGC 2-13-5 74.8 388 410550 22200 22219 GGTCCCCATGCTGGCCTCAG 5-10-5 69.4 389 410616 22200 22219 GGTCCCCATGCTGGCCTCAG 3-14-3 80.7 389 410695 22200 22219 GGTCCCCATGCTGGCCTCAG 2-13-5 85.5 389 410551 22201 22220 GGGTCCCCATGCTGGCCTCA 5-10-5 68.7 390 410696 22201 22220 GGGTCCCCATGCTGGCCTCA 2-13-5 86.3 390 410617 22201 22220 GGGTCCCCATGCTGGCCTCA 3-14-3 86.5 390 410552 22202 22221 CGGGTCCCCATGCTGGCCTC 5-10-5 77.9 391 410618 22202 22221 CGGGTCCCCATGCTGGCCTC 3-14-3 86.7 391 410697 22202 22221 CGGGTCCCCATGCTGGCCTC 2-13-5 87.0 391 410553 22203 22222 ACGGGTCCCCATGCTGGCCT 5-10-5 72.8 392 410619 22203 22222 ACGGGTCCCCATGCTGGCCT 3-14-3 84.1 392 410698 22203 22222 ACGGGTCCCCATGCTGGCCT 2-13-5 87.0 392 410699 22204 22223 CACGGGTCCCCATGCTGGCC 2-13-5 79.5 59 395185 22204 22223 CACGGGTCCCCATGCTGGCC 5-10-5 92.7 59 399907 22204 22223 CACGGGTCCCCATGCTGGCC 3-14-3 93.7 59 410620 22205 22224 ACACGGGTCCCCATGCTGGC 3-14-3 74.4 393 410554 22205 22224 ACACGGGTCCCCATGCTGGC 5-10-5 79.7 393 410700 22205 22224 ACACGGGTCCCCATGCTGGC 2-13-5 84.4 393 410621 22206 22225 GACACGGGTCCCCATGCTGG 3-14-3 76.0 394 410701 22206 22225 GACACGGGTCCCCATGCTGG 2-13-5 83.4 394 405991 22206 22225 GACACGGGTCCCCATGCTGG 5-10-5 91.9 394 410555 22207 22226 GGACACGGGTCCCCATGCTG 5-10-5 77.0 395 410622 22207 22226 GGACACGGGTCCCCATGCTG 3-14-3 79.8 395 410702 22207 22226 GGACACGGGTCCCCATGCTG 2-13-5 85.0 395 410703 22208 22227 TGGACACGGGTCCCCATGCT 2-13-5 70.4 396 410623 22208 22227 TGGACACGGGTCCCCATGCT 3-14-3 78.9 396 405992 22208 22227 TGGACACGGGTCCCCATGCT 5-10-5 89.3 396 410704 22209 22228 GTGGACACGGGTCCCCATGC 2-13-5 78.6 397 410556 22209 22228 GTGGACACGGGTCCCCATGC 5-10-5 81.4 397 410624 22209 22228 GTGGACACGGGTCCCCATGC 3-14-3 82.8 397 410625 22210 22229 AGTGGACACGGGTCCCCATG 3-14-3 64.4 398 410705 22210 22229 AGTGGACACGGGTCCCCATG 2-13-5 74.7 398 405993 22210 22229 AGTGGACACGGGTCCCCATG 5-10-5 85.5 398 410706 22211 22230 CAGTGGACACGGGTCCCCAT 2-13-5 70.8 399 410557 22211 22230 CAGTGGACACGGGTCCCCAT 5-10-5 74.2 399 410626 22211 22230 CAGTGGACACGGGTCCCCAT 3-14-3 76.8 399 410627 22212 22231 GCAGTGGACACGGGTCCCCA 3-14-3 71.4 400 410707 22212 22231 GCAGTGGACACGGGTCCCCA 2-13-5 72.8 400 405994 22212 22231 GCAGTGGACACGGGTCCCCA 5-10-5 95.2 400 410708 22213 22232 GGCAGTGGACACGGGTCCCC 2-13-5 79.0 401 410628 22213 22232 GGCAGTGGACACGGGTCCCC 3-14-3 88.1 401 410558 22213 22232 GGCAGTGGACACGGGTCCCC 5-10-5 88.9 401 410629 22214 22233 TGGCAGTGGACACGGGTCCC 3-14-3 66.5 402 410709 22214 22233 TGGCAGTGGACACGGGTCCC 2-13-5 66.9 402 405995 22214 22233 TGGCAGTGGACACGGGTCCC 5-10-5 92.4 402 410630 22215 22234 GTGGCAGTGGACACGGGTCC 3-14-3 54.1 403 410710 22215 22234 GTGGCAGTGGACACGGGTCC 2-13-5 61.4 403 410559 22215 22234 GTGGCAGTGGACACGGGTCC 5-10-5 77.5 403 410711 22216 22235 GGTGGCAGTGGACACGGGTC 2-13-5 NA 404 410631 22216 22235 GGTGGCAGTGGACACGGGTC 3-14-3 NA 404 410560 22216 22235 GGTGGCAGTGGACACGGGTC 5-10-5 42.5 404 410712 22217 22236 TGGTGGCAGTGGACACGGGT 2-13-5 NA 405 410632 22217 22236 TGGTGGCAGTGGACACGGGT 3-14-3 NA 405 410561 22217 22236 TGGTGGCAGTGGACACGGGT 5-10-5 NA 405 410774 22220 22239 TGTTGGTGGCAGTGGACACG 5-10-5 47.8 406 410776 23985 24004 GTGCCAAGGTCCTCCACCTC 5-10-5 77.3 408 410777 24005 24024 TCAGCACAGGCGGCTTGTGG 5-10-5 40.0 409 410778 24035 24054 CCACGCACTGGTTGGGCTGA 5-10-5 52.7 410 399908 24095 24114 CTTTGCATTCCAGACCTGGG 3-14-3 74.9 60 410713 24095 24114 CTTTGCATTCCAGACCTGGG 2-13-5 84.9 60 395186 24095 24114 CTTTGCATTCCAGACCTGGG 5-10-5 93.7 60 410714 24096 24115 ACTTTGCATTCCAGACCTGG 2-13-5 82.4 411 410633 24096 24115 ACTTTGCATTCCAGACCTGG 3-14-3 83.8 411 410562 24096 24115 ACTTTGCATTCCAGACCTGG 5-10-5 87.3 411 410715 24097 24116 GACTTTGCATTCCAGACCTG 2-13-5 80.5 412 410634 24097 24116 GACTTTGCATTCCAGACCTG 3-14-3 81.8 412 405996 24097 24116 GACTTTGCATTCCAGACCTG 5-10-5 91.0 412 410563 24098 24117 TGACTTTGCATTCCAGACCT 5-10-5 75.4 413 410635 24098 24117 TGACTTTGCATTCCAGACCT 3-14-3 75.9 413 410716 24098 24117 TGACTTTGCATTCCAGACCT 2-13-5 88.3 413 410636 24099 24118 TTGACTTTGCATTCCAGACC 3-14-3 61.3 414 410564 24099 24118 TTGACTTTGCATTCCAGACC 5-10-5 71.3 414 410717 24099 24118 TTGACTTTGCATTCCAGACC 2-13-5 76.7 414 410718 24100 24119 CTTGACTTTGCATTCCAGAC 2-13-5 55.8 61 399973 24100 24119 CTTGACTTTGCATTCCAGAC 3-14-3 82.1 61 399817 24100 24119 CTTGACTTTGCATTCCAGAC 5-10-5 93.6 61 405997 24102 24121 TCCTTGACTTTGCATTCCAG 5-10-5 95.6 415 410779 24115 24134 CGGGATTCCATGCTCCTTGA 5-10-5 79.7 416 410781 25994 26013 GGAGGGCACTGCAGCCAGTC 5-10-5 66.0 418 410719 26112 26131 CAGATGGCAACGGCTGTCAC 2-13-5 61.0 419 410637 26112 26131 CAGATGGCAACGGCTGTCAC 3-14-3 66.0 419 410565 26112 26131 CAGATGGCAACGGCTGTCAC 5-10-5 71.6 419 410638 26113 26132 GCAGATGGCAACGGCTGTCA 3-14-3 71.9 420 410720 26113 26132 GCAGATGGCAACGGCTGTCA 2-13-5 75.1 420 410566 26113 26132 GCAGATGGCAACGGCTGTCA 5-10-5 77.7 420 410639 26114 26133 AGCAGATGGCAACGGCTGTC 3-14-3 67.7 421 410721 26114 26133 AGCAGATGGCAACGGCTGTC 2-13-5 68.8 421 410567 26114 26133 AGCAGATGGCAACGGCTGTC 5-10-5 72.1 421 410568 26115 26134 CAGCAGATGGCAACGGCTGT 5-10-5 54.2 422 410640 26115 26134 CAGCAGATGGCAACGGCTGT 3-14-3 57.7 422 410722 26115 26134 CAGCAGATGGCAACGGCTGT 2-13-5 59.2 422 410569 26116 26135 GCAGCAGATGGCAACGGCTG 5-10-5 57.7 423 410641 26116 26135 GCAGCAGATGGCAACGGCTG 3-14-3 66.1 423 410723 26116 26135 GCAGCAGATGGCAACGGCTG 2-13-5 81.1 423 399909 26117 26136 GGCAGCAGATGGCAACGGCT 3-14-3 69.9 62 410724 26117 26136 GGCAGCAGATGGCAACGGCT 2-13-5 87.4 62 395187 26117 26136 GGCAGCAGATGGCAACGGCT 5-10-5 92.7 62 410642 26118 26137 CGGCAGCAGATGGCAACGGC 3-14-3 70.4 424 410725 26118 26137 CGGCAGCAGATGGCAACGGC 2-13-5 75.6 424 410725 26118 26137 CGGCAGCAGATGGCAACGGC 5-10-5 78.3 424 410571 26119 26138 CCGGCAGCAGATGGCAACGG 5-10-5 52.8 425 410643 26119 26138 CCGGCAGCAGATGGCAACGG 3-14-3 59.8 425 410726 26119 26138 CCGGCAGCAGATGGCAACGG 2-13-5 69.8 425 410644 26120 26139 TCCGGCAGCAGATGGCAACG 3-14-3 59.0 426 410727 26120 26139 TCCGGCAGCAGATGGCAACG 2-13-5 68.3 426 405998 26120 26139 TCCGGCAGCAGATGGCAACG 5-10-5 75.6 426 410572 26121 26140 CTCCGGCAGCAGATGGCAAC 5-10-5 40.9 427 410728 26121 26140 CTCCGGCAGCAGATGGCAAC 2-13-5 56.4 427 410645 26121 26140 CTCCGGCAGCAGATGGCAAC 3-14-3 70.2 427 410729 26122 26141 GCTCCGGCAGCAGATGGCAA 2-13-5 67.7 428 410573 26122 26141 GCTCCGGCAGCAGATGGCAA 5-10-5 71.8 428 410646 26122 26141 GCTCCGGCAGCAGATGGCAA 3-14-3 72.8 428 410782 26132 26151 CCAGGTGCCGGCTCCGGCAG 5-10-5 43.2 429 410783 26142 26161 GAGGCCTGCGCCAGGTGCCG 5-10-5 63.7 430 399819 26316 26335 CCCACTCAAGGGCCAGGCCA 5-10-5 93.4 65 399912 26404 26423 ATGCCCCACAGTGAGGGAGG 3-14-3 84.3 66 405893 26635 26654 ATGAGGGCCATCAGCACCTT 5-10-5 93.9 431 405894 26636 26655 GATGAGGGCCATCAGCACCT 5-10-5 93.9 432 405895 26637 26656 AGATGAGGGCCATCAGCACC 5-10-5 91.8 433 405896 26638 26657 GAGATGAGGGCCATCAGCAC 5-10-5 88.5 434 395194 26639 26658 GGAGATGAGGGCCATCAGCA 5-10-5 90.0 69 399916 26639 26658 GGAGATGAGGGCCATCAGCA 3-14-3 94.5 69 405897 26640 26659 TGGAGATGAGGGCCATCAGC 5-10-5 85.1 435 405898 26641 26660 CTGGAGATGAGGGCCATCAG 5-10-5 86.9 436 405899 26642 26661 GCTGGAGATGAGGGCCATCA 5-10-5 91.5 437 405900 26643 26662 AGCTGGAGATGAGGGCCATC 5-10-5 83.7 438 405901 26709 26728 GCTAGATGCCATCCAGAAAG 5-10-5 88.4 439 405902 26710 26729 GGCTAGATGCCATCCAGAAA 5-10-5 89.3 440 405903 26711 26730 TGGCTAGATGCCATCCAGAA 5-10-5 92.6 441 405904 26712 26731 CTGGCTAGATGCCATCCAGA 5-10-5 90.4 442 399821 26713 26732 TCTGGCTAGATGCCATCCAG 5-10-5 91.7 71 405905 26714 26733 CTCTGGCTAGATGCCATCCA 5-10-5 90.7 443 405906 26715 26734 CCTCTGGCTAGATGCCATCC 5-10-5 93.3 444 405907 26716 26735 GCCTCTGGCTAGATGCCATC 5-10-5 90.2 445 405908 26717 26736 AGCCTCTGGCTAGATGCCAT 5-10-5 83.3 446 399978 26795 26814 AGCCTGGCATAGAGCAGAGT 3-14-3 96.4 73

Antisense oligonucleotides that exhibited less than 30% inhibition of PCSK9 mRNA levels were marked with “NA”.

Antisense oligonucleotides with the following ISIS Nos exhibited at least 80% inhibition of PCSK9 mRNA levels: 395152, 395153, 395155, 395158, 395163, 395165, 395166, 395168, 395173, 395183, 395185, 395186, 395187, 395194, 399798, 399801, 399804, 399817, 399819, 399821, 399868, 399877, 399879, 399887, 399888, 399891, 399895, 399900, 399904, 399905, 399906, 399907, 399912, 399916, 399935, 399936, 399954, 399960, 399967, 399968, 399969, 399973, 399978, 405861, 405862, 405863, 405864, 405865, 405866, 405869, 405870, 405871, 405872, 405873, 405874, 405875, 405876, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405889, 405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405900, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405908, 405909, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921, 405922, 405923, 405924, 405925, 405926, 405927, 405928, 405929, 405930, 405931, 405932, 405934, 405935, 405936, 405940, 405941, 405943, 405944, 405949, 405952, 405953, 405954, 405955, 405956, 405958, 405959, 405960, 405961, 405973, 405974, 405976, 405977, 405978, 405979, 405980, 405982, 405984, 405987, 405988, 405989, 405990, 405991, 405992, 405993, 405994, 405995, 405996, 405997, 406003, 406005, 406008, 406009, 406014, 406016, 406020, 406023, 406026, 406028, 406029, 406030, 406031, 406032, 406033, 406035, 406036, 406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410530, 410556, 410558, 410562, 410578, 410583, 410589, 410590, 410591, 410592, 410593, 410595, 410609, 410611, 410616, 410617, 410618, 410619, 410624, 410628, 410633, 410634, 410647, 410648, 410650, 410652, 410658, 410664, 410666, 410667, 410668, 410671, 410677, 410686, 410688, 410689, 410693, 410695, 410696, 410697, 410698, 410700, 410701, 410702, 410713, 410714, 410715, 410716, 410723, 410724, 410730, 410754, 410757, 410759, 410769, and 410772. The target segments to which these antisense oligonucleotides are targeted are active target segments. The target regions to which these antisense oligonucleotides are targeted are active target regions.

ISIS Nos 395152, 395155, 395158, 395165, 395166, 395168, 395173, 395183, 395185, 395186, 395187, 395194, 399801, 399817, 399819, 399821, 399868, 399877, 399879, 399887, 399888, 399891, 399900, 399904, 399905, 399906, 399907, 399916, 399936, 399954, 399969, 399978, 405861, 405862, 405864, 405869, 405871, 405872, 405873, 405874, 405875, 405876, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921, 405922, 405923, 405924, 405925, 405926, 405927, 405928, 405929, 405930, 405931, 405932, 405934, 405935, 405936, 405941, 405952, 405953, 405954, 405955, 405956, 405959, 405960, 405961, 405974, 405977, 405978, 405979, 405980, 405982, 405987, 405988, 405989, 405990, 405991, 405992, 405993, 405994, 405995, 405996, 405997, 406008, 406009, 406014, 406016, 406023, 406028, 406029, 406030, 406031, 406032, 406033, 406035, 406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410558, 410562, 410583, 410591, 410592, 410593, 410611, 410617, 410618, 410628, 410647, 410650, 410668, 410677, 410688, 410695, 410696, 410697, 410698, 410702, 410716, 410724, 410730, 410757, and 410772 each exhibited at least 85% inhibition of PCSK9 mRNA levels.

ISIS Nos 395152, 395155, 395158, 395165, 395166, 395168, 395173, 395183, 395185, 395186, 395187, 399801, 399817, 399819, 399821, 399877, 399879, 399887, 399888, 399891, 399900, 399904, 399905, 399906, 399907, 399916, 399969, 399978, 405862, 405864, 405874, 405875, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405891, 405893, 405894, 405895, 405899, 405903, 405906, 405910, 405913, 405916, 405923, 405924, 405925, 405926, 405927, 405930, 405934, 405936, 405953, 405955, 405956, 405959, 405978, 405988, 405990, 405991, 405994, 405995, 405996, 405997, 406008, 406014, 406023, 406028, 406029, 406030, 406033, 406035, 406039, 406040, 406041, 406042, 406043, 406044, 410592, 410647, 410668, and 410772 each exhibited at least 90% inhibition of PCSK9 mRNA levels.

ISIS Nos 395165, 395166, 399801, 399978, 405881, 405891, 405959, 405978, 405988, 405990, 405994, 405997, 406008, 406030, 406040, 406041, and 406044 each exhibited at least 95% inhibition of PCSK9 mRNA levels.

TABLE 19 Antisense inhibition of PCSK9 in human cells (Hep3B) targeting SEQ ID NO: 3 5′ Target 3′ Target Site to Site to SEQ SEQ ID SEQ ID % ID Isis No. NO: 3 NO: 3 Sequence 5′-3′ Motif inhibition NO: 399935 1075 1094 AGGACCCAAGTCATCCTGCT 3-14-3 81.1 96 399936 1105 1124 GGCCATCAGCTGGCAATGCT 3-14-3 89.0 124 410753 1351 1370 ATACACCTCCACCAGGCTGC 5-10-5 72.8 215 406025 1362 1381 GTGTCTAGGAGATACACCTC 5-10-5 74.2 216 406026 1367 1386 TGCTGGTGTCTAGGAGATAC 5-10-5 80.5 217 405869 1392 1411 TCGATTTCCCGGTGGTCACT 5-10-5 87.6 218 405870 1393 1412 CTCGATTTCCCGGTGGTCAC 5-10-5 83.3 219 405871 1394 1413 CCTCGATTTCCCGGTGGTCA 5-10-5 87.0 220 405872 1395 1414 CCCTCGATTTCCCGGTGGTC 5-10-5 88.2 221 399798 1396 1415 GCCCTCGATTTCCCGGTGGT 5-10-5 84.3 22 399954 1396 1415 GCCCTCGATTTCCCGGTGGT 3-14-3 90.3 22 405873 1397 1416 TGCCCTCGATTTCCCGGTGG 5-10-5 90.0 222 405874 1398 1417 CTGCCCTCGATTTCCCGGTG 5-10-5 91.4 223 405875 1399 1418 CCTGCCCTCGATTTCCCGGT 5-10-5 93.5 224 405876 1400 1419 CCCTGCCCTCGATTTCCCGG 5-10-5 90.1 225 406027 1404 1423 ATGACCCTGCCCTCGATTTC 5-10-5 73.9 226 406028 1406 1425 CCATGACCCTGCCCTCGATT 5-10-5 92.3 227 406029 1408 1427 GACCATGACCCTGCCCTCGA 5-10-5 91.4 228 406030 1412 1431 CGGTGACCATGACCCTGCCC 5-10-5 95.6 229 406031 1414 1433 GTCGGTGACCATGACCCTGC 5-10-5 89.6 230 410733 1416 1435 AAGTCGGTGACCATGACCCT 5-10-5 65.7 231 406032 1418 1437 CGAAGTCGGTGACCATGACC 5-10-5 88.5 232 410754 1428 1447 GGCACATTCTCGAAGTCGGT 5-10-5 80.1 233 395163 1453 1472 GTGGAAGCGGGTCCCGTCCT 5-10-5 83.9 25 410755 1463 1482 TGGCCTGTCTGTGGAAGCGG 5-10-5 NA 234 410756 1490 1509 GGTGGGTGCCATGACTGTCA 5-10-5 67.1 235 406033 1497 1516 CCTGCCAGGTGGGTGCCATG 5-10-5 91.2 237 406034 1502 1521 CCACCCCTGCCAGGTGGGTG 5-10-5 59.3 238 406035 1507 1526 GCTGACCACCCCTGCCAGGT 5-10-5 91.7 239 410757 1515 1534 TCCCGGCCGCTGACCACCCC 5-10-5 88.9 240 406036 1519 1538 GGCATCCCGGCCGCTGACCA 5-10-5 84.0 241 406037 1522 1541 GCCGGCATCCCGGCCGCTGA 5-10-5 55.9 242 410536 1527 1546 GCCACGCCGGCATCCCGGCC 5-10-5 63.7 243 410658 1527 1546 GCCACGCCGGCATCCCGGCC 2-13-5 82.0 243 410583 1527 1546 GCCACGCCGGCATCCCGGCC 3-14-3 87.3 243 410537 1528 1547 GGCCACGCCGGCATCCCGGC 5-10-5 58.5 244 410659 1528 1547 GGCCACGCCGGCATCCCGGC 2-13-5 73.7 244 410584 1528 1547 GGCCACGCCGGCATCCCGGC 3-14-3 79.5 244 410660 1529 1548 TGGCCACGCCGGCATCCCGG 2-13-5 63.0 245 410585 1529 1548 TGGCCACGCCGGCATCCCGG 3-14-3 73.2 245 406038 1529 1548 TGGCCACGCCGGCATCCCGG 5-10-5 86.3 245 410661 1530 1549 TTGGCCACGCCGGCATCCCG 2-13-5 53.3 246 410586 1530 1549 TTGGCCACGCCGGCATCCCG 3-14-3 60.3 246 405877 1530 1549 TTGGCCACGCCGGCATCCCG 5-10-5 83.2 246 410587 1531 1550 CTTGGCCACGCCGGCATCCC 3-14-3 63.2 247 410662 1531 1550 CTTGGCCACGCCGGCATCCC 2-13-5 67.3 247 405878 1531 1550 CTTGGCCACGCCGGCATCCC 5-10-5 91.4 247 410588 1532 1551 CCTTGGCCACGCCGGCATCC 3-14-3 65.3 248 410663 1532 1551 CCTTGGCCACGCCGGCATCC 2-13-5 67.9 248 405879 1532 1551 CCTTGGCCACGCCGGCATCC 5-10-5 94.1 248 410589 1533 1552 CCCTTGGCCACGCCGGCATC 3-14-3 80.1 249 410664 1533 1552 CCCTTGGCCACGCCGGCATC 2-13-5 81.9 249 405880 1533 1552 CCCTTGGCCACGCCGGCATC 5-10-5 93.5 249 410665 1534 1553 ACCCTTGGCCACGCCGGCAT 2-13-5 78.7 28 399887 1534 1553 ACCCTTGGCCACGCCGGCAT 3-14-3 91.9 28 395165 1534 1553 ACCCTTGGCCACGCCGGCAT 5-10-5 96.7 28 410590 1535 1554 CACCCTTGGCCACGCCGGCA 3-14-3 82.2 250 410666 1535 1554 CACCCTTGGCCACGCCGGCA 2-13-5 82.7 250 405881 1535 1554 CACCCTTGGCCACGCCGGCA 5-10-5 98.3 250 410667 1536 1555 GCACCCTTGGCCACGCCGGC 2-13-5 84.7 251 410591 1536 1555 GCACCCTTGGCCACGCCGGC 3-14-3 86.3 251 405882 1536 1555 GCACCCTTGGCCACGCCGGC 5-10-5 91.6 251 410668 1537 1556 GGCACCCTTGGCCACGCCGG 2-13-5 91.4 252 405883 1537 1556 GGCACCCTTGGCCACGCCGG 5-10-5 92.5 252 410592 1537 1556 GGCACCCTTGGCCACGCCGG 3-14-3 93.4 252 410669 1538 1557 TGGCACCCTTGGCCACGCCG 2-13-5 69.0 253 410593 1538 1557 TGGCACCCTTGGCCACGCCG 3-14-3 88.8 253 405884 1538 1557 TGGCACCCTTGGCCACGCCG 5-10-5 90.8 253 410670 1539 1558 CTGGCACCCTTGGCCACGCC 2-13-5 74.9 254 410594 1539 1558 CTGGCACCCTTGGCCACGCC 3-14-3 75.2 254 410538 1539 1558 CTGGCACCCTTGGCCACGCC 5-10-5 78.2 254 410539 1540 1559 GCTGGCACCCTTGGCCACGC 5-10-5 61.9 255 410595 1540 1559 GCTGGCACCCTTGGCCACGC 3-14-3 83.1 255 410671 1540 1559 GCTGGCACCCTTGGCCACGC 2-13-5 83.3 255 410758 1545 1564 CGCATGCTGGCACCCTTGGC 5-10-5 73.6 256 406039 1566 1585 CAGTTGAGCACGCGCAGGCT 5-10-5 91.2 257 406040 1568 1587 GGCAGTTGAGCACGCGCAGG 5-10-5 96.5 258 399888 1570 1589 TTGGCAGTTGAGCACGCGCA 3-14-3 90.9 29 395166 1570 1589 TTGGCAGTTGAGCACGCGCA 5-10-5 96.2 29 406041 1572 1591 CCTTGGCAGTTGAGCACGCG 5-10-5 96.6 259 399801 1575 1594 TTCCCTTGGCAGTTGAGCAC 5-10-5 95.6 30 406042 1577 1596 CCTTCCCTTGGCAGTTGAGC 5-10-5 92.0 260 406043 1581 1600 GTGCCCTTCCCTTGGCAGTT 5-10-5 91.9 261 406044 1583 1602 CCGTGCCCTTCCCTTGGCAG 5-10-5 95.4 262 410759 1594 1613 GGTGCCGCTAACCGTGCCCT 5-10-5 83.7 263 410760 1618 1637 CCGAATAAACTCCAGGCCTA 5-10-5 96.7 265 406045 1626 1645 TGGCTTTTCCGAATAAACTC 5-10-5 88.4 266 395168 1628 1647 GCTGGCTTTTCCGAATAAAC 5-10-5 91.9 32 405909 1630 1649 CAGCTGGCTTTTCCGAATAA 5-10-5 80.3 267 405910 1632 1651 ACCAGCTGGCTTTTCCGAAT 5-10-5 90.9 268 405911 1634 1653 GGACCAGCTGGCTTTTCCGA 5-10-5 88.0 269 405912 1638 1657 GGCTGGACCAGCTGGCTTTT 5-10-5 78.2 270 410761 1649 1668 GTGGCCCCACAGGCTGGACC 5-10-5 53.7 271 410762 1662 1681 AGCAGCACCACCAGTGGCCC 5-10-5 57.6 272 410763 1684 1703 GCTGTACCCACCCGCCAGGG 5-10-5 68.8 273 410764 1730 1749 CGACCCCAGCCCTCGCCAGG 5-10-5 66.3 274 399891 1740 1759 GTGACCAGCACGACCCCAGC 3-14-3 91.1 33 405913 1742 1761 CGGTGACCAGCACGACCCCA 5-10-5 93.8 275 405914 1744 1763 AGCGGTGACCAGCACGACCC 5-10-5 90.4 276 405915 1746 1765 GCAGCGGTGACCAGCACGAC 5-10-5 86.8 277 405916 1748 1767 CGGCAGCGGTGACCAGCACG 5-10-5 91.3 278 410734 1749 1768 CCGGCAGCGGTGACCAGCAC 5-10-5 65.8 249 405917 1752 1771 TTGCCGGCAGCGGTGACCAG 5-10-5 62.0 280 405918 1754 1773 AGTTGCCGGCAGCGGTGACC 5-10-5 33.5 281 405919 1756 1775 GAAGTTGCCGGCAGCGGTGA 5-10-5 68.2 282 405920 1758 1777 CGGAAGTTGCCGGCAGCGGT 5-10-5 86.2 283 405921 1760 1779 CCCGGAAGTTGCCGGCAGCG 5-10-5 85.5 284 405922 1762 1781 GTCCCGGAAGTTGCCGGCAG 5-10-5 86.0 285 405937 1820 1839 GAGGCACCAATGATGTCCTC 5-10-5 77.7 306 405938 1824 1843 GCTGGAGGCACCAATGATGT 5-10-5 75.5 307 405939 1826 1845 TCGCTGGAGGCACCAATGAT 5-10-5 70.6 308 405940 1828 1847 AGTCGCTGGAGGCACCAATG 5-10-5 84.4 309 405941 1830 1849 GCAGTCGCTGGAGGCACCAA 5-10-5 90.1 310 399804 1833 1852 GCTGCAGTCGCTGGAGGCAC 5-10-5 80.8 40 399960 1833 1852 GCTGCAGTCGCTGGAGGCAC 3-14-3 82.4 40 405942 1835 1854 GTGCTGCAGTCGCTGGAGGC 5-10-5 69.8 311 405943 1837 1856 AGGTGCTGCAGTCGCTGGAG 5-10-5 83.6 312 410737 1839 1858 GCAGGTGCTGCAGTCGCTGG 5-10-5 49.8 313 405944 1840 1859 AGCAGGTGCTGCAGTCGCTG 5-10-5 80.7 314 405945 1842 1861 AAAGCAGGTGCTGCAGTCGC 5-10-5 41.1 315 405946 1846 1865 ACACAAAGCAGGTGCTGCAG 5-10-5 70.4 316 405947 1848 1867 TGACACAAAGCAGGTGCTGC 5-10-5 72.5 317 410769 1858 1877 TCCCACTCTGTGACACAAAG 5-10-5 84.9 318 405948 1900 1919 TCATGGCTGCAATGCCAGCC 5-10-5 45.9 319 399806 1903 1922 GCATCATGGCTGCAATGCCA 5-10-5 82.8 42 399962 1903 1922 GCATCATGGCTGCAATGCCA 3-14-3 86.1 42 405949 1905 1924 CAGCATCATGGCTGCAATGC 5-10-5 84.9 320 405950 1907 1926 GACAGCATCATGGCTGCAAT 5-10-5 76.9 321 405951 1909 1928 CAGACAGCATCATGGCTGCA 5-10-5 75.4 322 405952 1913 1932 TCGGCAGACAGCATCATGGC 5-10-5 85.1 323 410738 1915 1934 GCTCGGCAGACAGCATCATG 5-10-5 68.4 324 405953 1917 1936 CGGCTCGGCAGACAGCATCA 5-10-5 93.6 325 405954 1919 1938 TCCGGCTCGGCAGACAGCAT 5-10-5 87.5 326 410770 1933 1952 CGGCCAGGGTGAGCTCCGGC 5-10-5 68.5 327 405955 1946 1965 CTCTGCCTCAACTCGGCCAG 5-10-5 94.2 328 405956 1948 1967 GTCTCTGCCTCAACTCGGCC 5-10-5 94.3 329 405958 1952 1971 ATCAGTCTCTGCCTCAACTC 5-10-5 80.3 331 405959 1954 1973 GGATCAGTCTCTGCCTCAAC 5-10-5 95.4 332 405960 1956 1975 GTGGATCAGTCTCTGCCTCA 5-10-5 90.1 333 405961 1958 1977 AAGTGGATCAGTCTCTGCCT 5-10-5 88.8 334 405962 1961 1980 GAGAAGTGGATCAGTCTCTG 5-10-5 56.1 335 410739 1963 1982 CAGAGAAGTGGATCAGTCTC 5-10-5 NA 336 405963 1965 1984 GGCAGAGAAGTGGATCAGTC 5-10-5 59.7 337 405964 1969 1988 CTTTGGCAGAGAAGTGGATC 5-10-5 45.3 338 405965 1974 1993 GACATCTTTGGCAGAGAAGT 5-10-5 55.9 339 405966 1976 1995 ATGACATCTTTGGCAGAGAA 5-10-5 51.3 340 405967 1980 1999 ATTGATGACATCTTTGGCAG 5-10-5 64.4 341 405968 1982 2001 TCATTGATGACATCTTTGGC 5-10-5 74.6 342 399967 1985 2004 GCCTCATTGATGACATCTTT 3-14-3 80.1 48 405969 1987 2006 AGGCCTCATTGATGACATCT 5-10-5 63.0 343 405970 1989 2008 CCAGGCCTCATTGATGACAT 5-10-5 66.8 344 410740 1991 2010 AACCAGGCCTCATTGATGAC 5-10-5 46.8 345 405885 1993 2012 GGAACCAGGCCTCATTGATG 5-10-5 73.4 346 410596 1995 2014 AGGGAACCAGGCCTCATTGA 3-14-3 NA 348 410672 1995 2014 AGGGAACCAGGCCTCATTGA 2-13-5 NA 348 405887 1995 2014 AGGGAACCAGGCCTCATTGA 5-10-5 NA 348 410597 1996 2015 CAGGGAACCAGGCCTCATTG 3-14-3 NA 349 405888 1996 2015 CAGGGAACCAGGCCTCATTG 5-10-5 NA 349 410673 1996 2015 CAGGGAACCAGGCCTCATTG 2-13-5 NA 349 410674 1997 2016 TCAGGGAACCAGGCCTCATT 2-13-5 60.7 49 399812 1997 2016 TCAGGGAACCAGGCCTCATT 5-10-5 68.7 49 399968 1997 2016 TCAGGGAACCAGGCCTCATT 3-14-3 84.5 49 410598 1998 2017 CTCAGGGAACCAGGCCTCAT 3-14-3 74.5 350 410675 1998 2017 CTCAGGGAACCAGGCCTCAT 2-13-5 76.9 350 405889 1998 2017 CTCAGGGAACCAGGCCTCAT 5-10-5 80.2 350 410599 1999 2018 CCTCAGGGAACCAGGCCTCA 3-14-3 70.6 351 410676 1999 2018 CCTCAGGGAACCAGGCCTCA 2-13-5 77.8 351 405890 1999 2018 CCTCAGGGAACCAGGCCTCA 5-10-5 87.7 351 410600 2000 2019 TCCTCAGGGAACCAGGCCTC 3-14-3 76.5 352 410677 2000 2019 TCCTCAGGGAACCAGGCCTC 2-13-5 88.4 352 405891 2000 2019 TCCTCAGGGAACCAGGCCTC 5-10-5 96.1 352 405892 2001 2020 GTCCTCAGGGAACCAGGCCT 5-10-5 71.4 353 410601 2001 2020 GTCCTCAGGGAACCAGGCCT 3-14-3 72.7 353 410678 2001 2020 GTCCTCAGGGAACCAGGCCT 2-13-5 75.0 353 395178 2002 2021 GGTCCTCAGGGAACCAGGCC 5-10-5 76.7 50 410679 2002 2021 GGTCCTCAGGGAACCAGGCC 2-13-5 77.6 50 399900 2002 2021 GGTCCTCAGGGAACCAGGCC 3-14-3 92.0 50 408653 2003 2022 TGGTCCTCAGGGAACCAGGC 5-10-5 42.5 354 410602 2003 2022 TGGTCCTCAGGGAACCAGGC 3-14-3 66.5 354 410680 2003 2022 TGGTCCTCAGGGAACCAGGC 2-13-5 71.6 354 410603 2004 2023 CTGGTCCTCAGGGAACCAGG 3-14-3 40.8 355 410681 2004 2023 CTGGTCCTCAGGGAACCAGG 2-13-5 44.4 355 405971 2004 2023 CTGGTCCTCAGGGAACCAGG 5-10-5 65.5 355 410540 2005 2024 GCTGGTCCTCAGGGAACCAG 5-10-5 50.9 356 410682 2005 2024 GCTGGTCCTCAGGGAACCAG 2-13-5 54.1 356 410604 2005 2024 GCTGGTCCTCAGGGAACCAG 3-14-3 62.5 356 405972 2006 2025 CGCTGGTCCTCAGGGAACCA 5-10-5 77.6 357 405973 2011 2030 GTACCCGCTGGTCCTCAGGG 5-10-5 83.8 358 405974 2013 2032 CAGTACCCGCTGGTCCTCAG 5-10-5 89.0 359 399813 2016 2035 GGTCAGTACCCGCTGGTCCT 5-10-5 72.4 51 399969 2016 2035 GGTCAGTACCCGCTGGTCCT 3-14-3 93.4 51 410771 2061 2080 ACCTGCCCCATGGGTGCTGG 5-10-5 NA 360 395181 2073 2092 AAACAGCTGCCAACCTGCCC 5-10-5 87.5 52 405975 2075 2094 CAAAACAGCTGCCAACCTGC 5-10-5 77.4 361 405976 2080 2099 TCCTGCAAAACAGCTGCCAA 5-10-5 81.2 362 405977 2082 2101 AGTCCTGCAAAACAGCTGCC 5-10-5 89.5 363 399992 2095 2114 GTGCTGACCACACAGTCCTG 3-14-3 92.6 130 405978 2105 2124 GGCCCCGAGTGTGCTGACCA 5-10-5 95.3 365 399904 2108 2127 GTAGGCCCCGAGTGTGCTGA 3-14-3 92.6 54 405979 2110 2129 GTGTAGGCCCCGAGTGTGCT 5-10-5 85.5 366 405980 2112 2131 CCGTGTAGGCCCCGAGTGTG 5-10-5 86.5 367 405981 2114 2133 ATCCGTGTAGGCCCCGAGTG 5-10-5 77.8 368 410741 2116 2135 CCATCCGTGTAGGCCCCGAG 5-10-5 78.4 369 405982 2118 2137 GGCCATCCGTGTAGGCCCCG 5-10-5 86.7 370 405983 2120 2139 GTGGCCATCCGTGTAGGCCC 5-10-5 73.1 371 405984 2168 2187 CTGGAGCAGCTCAGCAGCTC 5-10-5 83.4 372 405985 2170 2189 AACTGGAGCAGCTCAGCAGC 5-10-5 50.7 373 405986 2175 2194 GGAGAAACTGGAGCAGCTCA 5-10-5 75.6 374 405987 2177 2196 CTGGAGAAACTGGAGCAGCT 5-10-5 88.0 375 410776 2245 2264 GTGCCAAGGTCCTCCACCTC 5-10-5 77.3 408 410777 2265 2284 TCAGCACAGGCGGCTTGTGG 5-10-5 40.0 409 410778 2295 2314 CCACGCACTGGTTGGGCTGA 5-10-5 52.7 410 399908 2355 2374 CTTTGCATTCCAGACCTGGG 3-14-3 74.9 60 410713 2355 2374 CTTTGCATTCCAGACCTGGG 2-13-5 84.9 60 395186 2355 2374 CTTTGCATTCCAGACCTGGG 5-10-5 93.7 60 410714 2356 2375 ACTTTGCATTCCAGACCTGG 2-13-5 82.4 411 410633 2356 2375 ACTTTGCATTCCAGACCTGG 3-14-3 83.8 411 410562 2356 2375 ACTTTGCATTCCAGACCTGG 5-10-5 87.3 411 410715 2357 2376 GACTTTGCATTCCAGACCTG 2-13-5 80.5 412 410634 2357 2376 GACTTTGCATTCCAGACCTG 3-14-3 81.8 412 405996 2357 2376 GACTTTGCATTCCAGACCTG 5-10-5 91.0 412 410563 2358 2377 TGACTTTGCATTCCAGACCT 5-10-5 75.4 413 410635 2358 2377 TGACTTTGCATTCCAGACCT 3-14-3 75.9 413 410716 2358 2377 TGACTTTGCATTCCAGACCT 2-13-5 88.3 413 410636 2359 2378 TTGACTTTGCATTCCAGACC 3-14-3 61.3 414 410564 2359 2378 TTGACTTTGCATTCCAGACC 5-10-5 71.3 414 410717 2359 2378 TTGACTTTGCATTCCAGACC 2-13-5 76.7 414 410718 2360 2379 CTTGACTTTGCATTCCAGAC 2-13-5 55.8 61 399973 2360 2379 CTTGACTTTGCATTCCAGAC 3-14-3 82.1 61 399817 2360 2379 CTTGACTTTGCATTCCAGAC 5-10-5 93.6 61 405997 2362 2381 TCCTTGACTTTGCATTCCAG 5-10-5 95.6 415 410779 2375 2394 CGGGATTCCATGCTCCTTGA 5-10-5 79.7 416 410780 2405 2424 GCAGGCCACGGTCACCTGCT 5-10-5 60.9 417 410781 2442 2461 GGAGGGCACTGCAGCCAGTC 5-10-5 66.0 418 410719 2560 2579 CAGATGGCAACGGCTGTCAC 2-13-5 61.0 419 410637 2560 2579 CAGATGGCAACGGCTGTCAC 3-14-3 66.0 419 410565 2560 2579 CAGATGGCAACGGCTGTCAC 5-10-5 71.6 419 410638 2561 2580 GCAGATGGCAACGGCTGTCA 3-14-3 71.9 420 410720 2561 2580 GCAGATGGCAACGGCTGTCA 2-13-5 75.1 420 410566 2561 2580 GCAGATGGCAACGGCTGTCA 5-10-5 77.7 420 410639 2562 2581 AGCAGATGGCAACGGCTGTC 3-14-3 67.7 421 410721 2562 2581 AGCAGATGGCAACGGCTGTC 2-13-5 68.8 421 410567 2562 2581 AGCAGATGGCAACGGCTGTC 5-10-5 72.1 421 410568 2563 2582 CAGCAGATGGCAACGGCTGT 5-10-5 54.2 422 410640 2563 2582 CAGCAGATGGCAACGGCTGT 3-14-3 57.7 422 410722 2563 2582 CAGCAGATGGCAACGGCTGT 2-13-5 59.2 422 410569 2564 2583 GCAGCAGATGGCAACGGCTG 5-10-5 57.7 423 410641 2564 2583 GCAGCAGATGGCAACGGCTG 3-14-3 66.1 423 410723 2564 2583 GCAGCAGATGGCAACGGCTG 2-13-5 81.1 423 399909 2565 2584 GGCAGCAGATGGCAACGGCT 3-14-3 69.9 62 410724 2565 2584 GGCAGCAGATGGCAACGGCT 2-13-5 87.4 62 395187 2565 2584 GGCAGCAGATGGCAACGGCT 5-10-5 92.7 62 410642 2566 2585 CGGCAGCAGATGGCAACGGC 3-14-3 70.4 424 410725 2566 2585 CGGCAGCAGATGGCAACGGC 2-13-5 75.6 424 410570 2566 2585 CGGCAGCAGATGGCAACGGC 5-10-5 78.3 424 410571 2567 2586 CCGGCAGCAGATGGCAACGG 5-10-5 52.8 425 410643 2567 2586 CCGGCAGCAGATGGCAACGG 3-14-3 59.8 425 410726 2567 2586 CCGGCAGCAGATGGCAACGG 2-13-5 69.8 425 410644 2568 2587 TCCGGCAGCAGATGGCAACG 3-14-3 59.0 426 410727 2568 2587 TCCGGCAGCAGATGGCAACG 2-13-5 68.3 426 405998 2568 2587 TCCGGCAGCAGATGGCAACG 5-10-5 75.6 426 405988 2568 2587 TCCGGCAGCAGATGGCAACG 5-10-5 84.5 426 410572 2569 2588 CTCCGGCAGCAGATGGCAAC 5-10-5 40.9 427 410728 2569 2588 CTCCGGCAGCAGATGGCAAC 2-13-5 56.4 427 410645 2569 2588 CTCCGGCAGCAGATGGCAAC 3-14-3 70.2 427 410729 2570 2589 GCTCCGGCAGCAGATGGCAA 2-13-5 67.7 428 410573 2570 2589 GCTCCGGCAGCAGATGGCAA 5-10-5 71.8 428 410646 2570 2589 GCTCCGGCAGCAGATGGCAA 3-14-3 72.8 428 410782 2580 2599 CCAGGTGCCGGCTCCGGCAG 5-10-5 43.2 429 410783 2590 2609 GAGGCCTGCGCCAGGTGCCG 5-10-5 63.7 430 399819 2764 2783 CCCACTCAAGGGCCAGGCCA 5-10-5 93.4 65 399912 2852 2871 ATGCCCCACAGTGAGGGAGG 3-14-3 84.3 66 405893 3083 3102 ATGAGGGCCATCAGCACCTT 5-10-5 93.9 431 405894 3084 3103 GATGAGGGCCATCAGCACCT 5-10-5 93.9 432 405895 3085 3104 AGATGAGGGCCATCAGCACC 5-10-5 91.8 433 405896 3086 3105 GAGATGAGGGCCATCAGCAC 5-10-5 88.5 434 395194 3087 3106 GGAGATGAGGGCCATCAGCA 5-10-5 90.0 69 399916 3087 3106 GGAGATGAGGGCCATCAGCA 3-14-3 94.5 69 405897 3088 3107 TGGAGATGAGGGCCATCAGC 5-10-5 85.1 435 405898 3089 3108 CTGGAGATGAGGGCCATCAG 5-10-5 86.9 436 405899 3090 3109 GCTGGAGATGAGGGCCATCA 5-10-5 91.5 437 405900 3091 3110 AGCTGGAGATGAGGGCCATC 5-10-5 83.7 438 405901 3157 3176 GCTAGATGCCATCCAGAAAG 5-10-5 88.4 439 405902 3158 3177 GGCTAGATGCCATCCAGAAA 5-10-5 89.3 440 405903 3159 3178 TGGCTAGATGCCATCCAGAA 5-10-5 92.6 441 405904 3160 3179 CTGGCTAGATGCCATCCAGA 5-10-5 90.4 442 399821 3161 3180 TCTGGCTAGATGCCATCCAG 5-10-5 91.7 71 405905 3162 3181 CTCTGGCTAGATGCCATCCA 5-10-5 90.7 443 405906 3163 3182 CCTCTGGCTAGATGCCATCC 5-10-5 93.3 444 405907 3164 3183 GCCTCTGGCTAGATGCCATC 5-10-5 90.2 445 405908 3165 3184 AGCCTCTGGCTAGATGCCAT 5-10-5 83.3 446 399978 3243 3262 AGCCTGGCATAGAGCAGAGT 3-14-3 96.4 73

Antisense oligonucleotides that exhibited less than 30% inhibition of PCSK9 mRNA levels were marked with “NA”.

Antisense oligonucleotides with the following ISIS Nos exhibited at least 80% inhibition of PCSK9 mRNA levels: 395163, 395165, 395166, 395168, 395181, 395186, 395187, 395194, 399798, 399801, 399804, 399806, 399817, 399819, 399821, 399887, 399888, 399891, 399900, 399904, 399912, 399916, 399935, 399936, 399954, 399960, 399962, 399967, 399968, 399969, 399973, 399978, 399992, 405869, 405870, 405871, 405872, 405873, 405874, 405875, 405876, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405889, 405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405900, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405908, 405909, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921, 405922, 405940, 405941, 405943, 405944, 405949, 405952, 405953, 405954, 405955, 405956, 405958, 405959, 405960, 405961, 405973, 405974, 405976, 405977, 405978, 405979, 405980, 405982, 405984, 405987, 405988, 405996, 405997, 406026, 406028, 406029, 406030, 406031, 406032, 406033, 406035, 406036, 406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410562, 410583, 410589, 410590, 410591, 410592, 410593, 410595, 410633, 410634, 410658, 410664, 410666, 410667, 410668, 410671, 410677, 410713, 410714, 410715, 410716, 410723, 410724, 410754, 410757, 410759, 410760, and 410769. The target segments to which these antisense oligonucleotides are targeted are active target segments. The target regions to which these antisense oligonucleotides are targeted are active target regions.

ISIS Nos 395165, 395166, 395168, 395181, 395186, 395187, 395194, 399801, 399817, 399819, 399821, 399887, 399888, 399891, 399900, 399904, 399916, 399936, 399954, 399962, 399969, 399978, 399992, 405869, 405871, 405872, 405873, 405874, 405875, 405876, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921, 405922, 405941, 405952, 405953, 405954, 405955, 405956, 405959, 405960, 405961, 405974, 405977, 405978, 405979, 405980, 405982, 405987, 405996, 405997, 406028, 406029, 406030, 406031, 406032, 406033, 406035, 406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410562, 410583, 410591, 410592, 410593, 410668, 410677, 410716, 410724, 410757, and 410760 each exhibited at least 85% inhibition of PCSK9 mRNA levels.

ISIS Nos 395165, 395166, 395168, 395186, 395187, 395194, 399801, 399817, 399819, 399821, 399887, 399888, 399891, 399900, 399904, 399916, 399954, 399969, 399978, 399992, 405873, 405874, 405875, 405876, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405891, 405893, 405894, 405895, 405899, 405903, 405904, 405905, 405906, 405907, 405910, 405913, 405914, 405916, 405941, 405953, 405955, 405956, 405959, 405960, 405978, 405996, 405997, 406028, 406029, 406030, 406033, 406035, 406039, 406040, 406041, 406042, 406043, 406044, 410592, 410668, and 410760 each exhibited at least 90% inhibition of PCSK9 mRNA levels.

ISIS Nos 395165, 395166, 399801, 399978, 405881, 405891, 405959, 405978, 405997, 406030, 406040, 406041, 406044, and 410760 each exhibited at least 95% inhibition of PCSK9 mRNA levels.

Example 4 Antisense Reduction of Human PCSK9 mRNA in HepG2 Cells: Dose Response Experiment

Antisense oligonucleotides targeted to PCSK9 were tested at various doses in HepG2 cells. Cells were plated at densities of 10,000 cells per well and treated with NM concentrations of antisense oligonucleotide as indicated in Tables 20 and 21. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. Two different human PCSK9 primer probe sets were used to measure mRNA levels. Results with Primer Probe Set 2740 are shown in Table 20, and mRNA levels measured using primer probe set 2823 are shown in Table 21. Results are presented as percent PCSK9 mRNA levels relative to a control, as adjusted according to total RNA content measured by RIBOGREEN®. As illustrated in Tables 20 and 21, PCSK9 mRNA levels were reduced in a dose-dependent manner.

TABLE 20 Antisense Reduction of PCSK9 mRNA in HepG2 cells, Primer Probe Set 2740 6.25 200.0 Isis No nM 12.5 nM 25.0 nM 50.0 nM 100.0 nM nM 399819 93 113 84 71 26 12 395185 81 64 50 35 24 13 399916 65 56 41 18 20 25 399907 68 66 42 26 30 25 399954 71 63 38 22 16 15 395165 71 67 47 29 21 10 399936 65 51 31 20 30 28 399793 61 57 44 31 39 15 399969 57 54 40 27 25 19 395152 75 58 58 49 24 28

TABLE 21 Antisense Reduction of PCSK9 mRNA in HepG2 cells, Primer Probe Set 2823 6.25 200.0 Isis No nM 12.5 nM 25.0 nM 50.0 nM 100.0 nM nM 399819 80 264 87 85 34 15 395185 86 66 57 45 29 22 399916 75 47 45 20 30 45 399907 67 72 47 29 31 32 399954 59 50 35 17 22 27 395165 74 54 41 37 26 22 399936 62 41 33 32 28 42 399793 62 51 38 36 44 27 399969 73 40 58 28 29 35 395152 78 53 64 49 22 35

Example 5 Antisense Reduction of Human PCSK9 mRNA in Hep3B Cells: Dose Response Experiment

Antisense oligonucleotides targeted to PCSK9 were tested at various doses in Hep3B cells. Cells were plated at densities of 4,500 cells per well and treated with nM concentrations of antisense oligonucleotide as indicated in Table 22. ISIS 141923 is not complementary to any known gene sequence and is used in this and the following experiments as a negative control. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. Two different human PCSK9 primer probe sets were used to measure mRNA levels. Results with Primer Probe Set 2740 are shown in Table 22. Results are presented as percent PCSK9 mRNA levels relative to a control, as adjusted according to total RNA content measured by RIBOGREEN®. As illustrated in Table 22, PCSK9 mRNA levels were reduced in a dose-dependent manner.

TABLE 22 Antisense Reduction of PCSK9 mRNA in Hep3B cells, Primer Probe Set 2740 Isis No. 2.78 nM 8.33 nM 25.0 nM 75.0 nM 399819 127.5 85.0 36.1 21.9 405891 133.7 111.4 40.2 27.2 406008 144.6 108.3 49.9 17.7 395186 124.4 106.4 42.2 30.7 395185 150.7 106.6 53.6 38.1 405994 141.9 101.2 49.3 30.8 405988 148.1 117.4 47.8 25.2 406033 138.1 109.9 62.4 35.3 395187 133.0 115.6 59.9 31.6 405995 124.1 109.2 57.8 42.9 406023 130.0 103.5 67.1 28.0 399900 131.2 95.7 81.1 30.9 301012 121.2 100.1 63.8 49.6 395165 113.2 86.9 41.6 13.3 405879 124.6 88.8 44.1 13.1 405991 100.6 101.4 95.9 41.0 405923 130.4 115.0 68.9 35.1 395152 143.8 98.6 61.3 59.0 405881 100.7 77.4 50.2 4.5 141923 131.4 131.9 144.0 167.3

Example 6 Antisense Reduction of Human PCSK9 mRNA in HeLa Cells: Dose Response Experiment

Antisense oligonucleotides targeted to PCSK9 were tested at various doses in HeLa cells. Cells were plated at densities of 5,000 cells per well and treated with nM concentrations of antisense oligonucleotide as indicated in Table 23. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. Two different human PCSK9 primer probe sets were used to measure mRNA levels. Results with Primer Probe Set 2740 are shown in Table 23. Results are presented as percent PCSK9 mRNA levels relative to a control, as adjusted according to total RNA content measured by RIBOGREEN®. As illustrated in Table 23, PCSK9 mRNA levels were reduced in a dose-dependent manner.

TABLE 23 Antisense Reduction of PCSK9 mRNA in HeLa cells, Primer Probe Set 2740 Isis No. 2.963 nM 8.8889 nM 26.6667 nM 80.0 nM 399819 57 32 16 14 405891 57 46 17 18 406008 45 27 11 5.3 395186 40 28 11 6.6 395185 53 32 21 17 405994 48 34 20 13 405988 44 32 12 7.3 406033 61 42 25 12 395187 42 35 12 6.6 405995 54 39 21 15 406023 76 46 25 6.2 399900 85 48 23 6.4 301012 121 109 76 65 395165 61 30 10 2 405879 64 32 9.7 1 405991 64 35 24 10 405923 71 45 22 5 395152 73 34 13 3.1 405881 58 33 19 2.8 141923 122 118 131 140

Example 7 Antisense Reduction of Human PCSK9 mRNA in Primary Hepatocytes

Antisense oligonucleotides targeted to PCSK9 were tested for antisense inhibition of PCSK9 in human primary hepatocytes. Human primary hepatocytes were purchased from a commercial supplier, and cultured according to routine culture procedures. Cells were treated with 10, 25, 50, 150, or 300 nM of antisense oligonucleotide for a period of 24 hours, after which RNA was isolated and PCSK9 mRNA was measured by real-time PCR as described herein.

The antisense oligonucleotides tested in human primary hepatocytes were Isis 395152, 395155, 395165, 395185, 399819, 399821, 399916, 399954, 399978, and 399992. The antisense oligonucleotides inhibited PCSK9 in a dose-dependent manner in human primary hepatocytes.

Monkey primary hepatocytes were similarly treated with 10, 25, 50, 150, or 300 nM concentrations of antisense oligonucleotide targeted to PCSK9. The antisense oligonucleotides tested in monkey primary hepatocytes were Isis 395152, 395155, 395165, 395185, 399819, 399821, 399916, 399954, 399978, and 399992. Each of these antisense oligonucleotides is fully complementary to a monkey PCSK9 nucleic acid. The antisense oligonucleotides inhibited PCSK9 in a dose-dependent manner in monkey primary hepatocytes.

Example 8 Additional Antisense Reduction of Human PCSK9 mRNA in Primary Hepatocytes

Antisense oligonucleotides targeted to PCSK9 were tested for antisense inhibition of PCSK9 in human primary hepatocytes. Human primary hepatocytes were purchased from a commercial supplier, and cultured according to routine culture procedures. Cells were treated with 10, 25, 50, 150, or 300 nM of antisense oligonucleotide for a period of 24 hours, after which RNA was isolated and PCSK9 mRNA was measured by real-time PCR as described herein.

The antisense oligonucleotides tested in human primary hepatocytes were Isis 395165, 395185, 395186, 395187, 405879, 405881, 405891, 405988, 405994, and 406008. The antisense oligonucleotides inhibited PCSK9 in a dose-dependent manner in human primary hepatocytes as shown in Table 24.

TABLE 24 Dose-dependent Reduction in Human PCSK9 mRNA Expression in Human Primary Hepatocytes ISIS No. 10 nM 25 nM 50 nM 150 nM 300 nM 395165 76 48.7 22.5 20.2 16.2 395185 74.5 42.9 52.6 25.5 15.7 395186 123.8 58.3 34.4 21 19.9 395187 98.7 43.5 29.7 17.4 21.1 405879 119.2 176.6 64.9 48.4 47.7 405881 222.4 115.3 49.5 22.3 79.3 405891 169.9 138.7 73.9 86.9 48.7 405988 85.2 105.4 67.8 43.9 43.8 405994 95.7 83.3 43.5 24.6 10.5 406008 155 101.7 116 41.7 32.4

Example 9 Antisense Reduction of PCSK9 mRNA in Cyno Primary Hepatocytes

Antisense oligonucleotides targeted to PCSK9 were tested for antisense inhibition of PCSK9 in cyno primary hepatocytes. Cyno primary hepatocytes were purchased from a commercial supplier, and cultured according to routine culture procedures. Cells were treated with 10, 25, 50, 150, or 300 nM of antisense oligonucleotide for a period of 24 hours, after which RNA was isolated and PCSK9 mRNA was measured by real-time PCR as described herein.

The antisense oligonucleotides tested in cyno primary hepatocytes were Isis 395165, 395185, 395186, 395187, 405879, 405881, 405891, 405988, 405994, and 406008. The antisense oligonucleotides inhibited PCSK9 in a dose-dependent manner in cyno primary hepatocytes as shown in Table 25.

TABLE 25 Dose-dependent PCSK9 mRNA Inhibition in Cyno Primary Hepatocytes ISIS No. 10 nM 25 nM 50 nM 150 nM 300 nM 395165 36.3 29.9 14.9 8.6 3.2 395185 65.0 31.9 35.1 16.6 12.0 395186 65.8 42.3 21.6 23.3 16.7 395187 61.3 32.9 13.2 7.7 9.02 405879 24.3 21.9 7.7 7.1 3.0 405881 75.0 33.5 26.5 11.2 4.9 405891 52.4 24.8 11.2 10.8 6.7 405988 59.4 23.7 22.5 10.7 8.2 405994 64.8 52.7 22.2 7.9 6.5 406008 56.3 28.2 26.5 13.4 13.3

Example 10 Antisense Reduction of Mouse PCSK9 mRNA In Vitro

Antisense oligonucleotides were designed to target murine PCSK9, and were evaluated for their ability to reduce PCSK9 mRNA in primary mouse hepatocytes.

Primary mouse hepatocytes were treated with antisense oligonucleotides at doses of 50, 150, or 300 nM, for a period of 24 hours. RNA was isolated using QIAGEN® RNeasy isolation kits and subjected to quantitative real-time PCR using commercially available reagents (Invitrogen, Carlsbad, Calif.). PCSK9 mRNA levels were measured using a mouse PCSK9 primer probe set, and normalized to G3PDH mRNA levels and/or RIBOGREEN® levels.

ISIS 394814 (GAGCAACTTCGGAGGCAGC, SEQ ID NO: 456) was identified as a potent inhibitor of mouse PCSK9. PCSK9 mRNA levels ISIS 394814 yielded a 50% reduction in PCSK9 mRNA at a concentration of 25 nM (i.e., an IC₅₀ of 25 nM) after a 24 hour incubation with primary mouse hepatocytes. No effect on cultured cell viability was observed.

Example 11 Antisense Reduction of PCSK9 mRNA in an Animal Model of Hyperlipidemia: High Fat Fed Mice

Treatment

C57BL/6 mice fed a high-fat diet are routinely employed as an animal model of hyperlipidemia, as well as an animal model of atherosclerosis. Accordingly, to evaluate the effects of PCSK9 antisense inhibition on serum lipid levels in vivo, ISIS 394814 was evaluated in C57BL/6 mice fed a high-fat diet. C57BL/6 mice 4-5 weeks of age were obtained from the Jackson Laboratory and maintained on a diet consisting of 60% fat. Treatment groups of 5 mice each were as follows: a group treated with ISIS 394814; a control group treated with saline; and control group treated with an oligonucleotide not complementary to any known gene sequence (ISIS 141923). Oligonucleotide or saline was administered intraperitoneally twice weekly, for a period of 6 weeks; oligonucleotide doses were 50 mg/kg. After the treatment period, whole blood was collected for analysis of serum parameters, and whole liver was collected for RNA analysis, protein analysis, and histological evaluation. Statistical analyses included a nonparametric, two-tailed t-test comparison of experimental samples (ISIS 394814) to control samples (saline or control oligonucleotide).

RNA Analysis

Liver RNA was isolated for real-time PCR analysis of PCSK9, LDL-receptor and apolipoprotein B mRNA levels. Treatment with ISIS 394814 resulted in a 92% reduction in PCSK9 mRNA levels, while no significant reduction in PCSK9 mRNA levels was observed in ISIS 141923-treated or saline-treated mice. Antisense inhibition of PCSK9 expression did not significantly affect liver LDL-receptor or liver apolipoprotein B mRNA levels.

Protein Analysis

Immunoblotting was performed to assess the effects of PCSK9 antisense inhibition on LDL-receptor protein and apolipoprotein B levels. Protein isolated from mouse liver was subjected to electrophoresis, transferred to a polyvinylidene-fluoride membrane, and subsequently probed with an anti-mouse LDL-receptor antibody and a scavenger receptor B1 (SR-B1) antibody. Mouse plasma was similarly subjected to electrophoresis and immunoblotting using an anti-mouse apolipoprotein B antibody. Secondary antibodies conjugated to peroxidase were used to detect primary antibodies. Protein bands were visualized using the ECL plus Western blot detection kit (Amersham Biosciences, UK) and quantified using ImageQuant™ analysis software (MolecularDynamics, Santa Clara, Calif.).

While LDL-receptor mRNA levels were not significantly affected, antisense inhibition of PCSK9 resulted in an approximate 2-fold increase in the level of hepatic LDL-receptor protein relative to 141923-treated controls. No changes in SR-B1 protein were observed. Furthermore, serum apoB-100 levels were significantly reduced by 50%, relative to 141923-treated controls. Serum apoB-48 levels were significantly increased by approximately 3-fold relative to 141923-treated controls. No significant changes in apoA-I protein were observed, relative to 141923-treated controls.

The mRNA levels of the RNA editing enzyme apobec-1 were also measured. Apobec-1 is responsible for the RNA editing that produces apoB-48 in murine liver and intestine. Human apobec-1 is expressed only in intestinal cells, thus these cells are the source of apoB-48 in humans. Antisense inhibition of PCSK9 resulted in an increase in murine hepatic apobec-1 mRNA levels by approximately 2.7 fold compared to saline-treated controls

Serum Lipid Analysis

Plasma concentrations of total cholesterol, LDL-C, HDL-C, free cholesterol, triglycerides, glucose, ketones, transaminases, and phospholipids were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, N.Y.). Serum lipoprotein and cholesterol profiling was performed as described by Crooke et al. (Lipid Res., 2005, 46, 872-884) using a Beckman System Gold 126 HPLC system, a 507e refrigerated autosampler, a 126 photodiode array detector (Beckman Instruments, Fullerton, Calif.), and a Superose 6 HR 10/30 column (Pfizer, Chicago, Ill.). HDL, LDL and VLDL fractions were measured at a wavelength of 505 nm and validated with a cholesterol calibration kit. (Sigma).

Administration of ISIS 394814 resulted in a 52% reduction in total cholesterol (saline, 183 mg/dL±18; ISIS 141923, 194 mg/dL±14; ISIS 394814, 87 mg/dL±19, p<0.005) and a 36% reduction in LDL-C (saline 22 mg/dL±4; ISIS 141923, 25 mg/dL±2; ISIS 394814, 14 mg/dL±2, p<0.005). Serum free cholesterol was reduced by 25% (saline 41 mg/dL±10; ISIS 141923, 58 mg/dL±3; ISIS 394814, 31 mg/dL±5, p<0.005) and phospholipids were reduced 54% (saline, 354 mg/dL±29; ISIS 141923, 383 mg/dL±21; ISIS 394814, 169 mg/dL±35, p<0.0001). HDL-C was also reduced by approximately 54% (saline, 183 mg/dL±18; ISIS 141923, 194 mg/dL±14; ISIS 394814, 87 mg/dL±19; p<0.0001). HPLC profiling confirmed the reduction of LDL and HDL lipid classes.

Liver Triglyceride Analysis

Liver triglyceride content was measured according to routine experimental procedures, for example, per procedures described by Desai et al., Diabetes, 2001, 50:2287-2295.

Antisense inhibition of PCSK9 for 6 weeks reduced liver triglyceride content by approximately 65% (p=0.01) relative to saline controls. No statistically significant changes in liver triglyceride content following 141923 treatment were observed.

Accordingly, one embodiment is a method of lowering LDL-C levels through the administration of an antisense oligonucleotide targeted to a PCSK9 nucleic acid. An additional embodiment includes a method of lowering total cholesterol through the administration of an antisense oligonucleotide targeted to a PCSK9 nucleic acid. An addition embodiment is lowering liver triglycerides by administering an antisense oligonucleotide targeted to a PCSK9 target nucleic acid.

Example 12 Antisense Reduction of PCSK9 mRNA in LDL-R Deficient/apoB-100 Animals

LDL-receptor (LDL-R)-deficient/apoB-100 animals do not express the LDL-R gene, and express only the apoB-100 form of apoB. PCSK9 antisense oligonucleotide was administered to LDL-R deficient/apoB-100 mice, to evaluate the effects of antisense inhibition of PCSK9 in the absence of the LDL-R. Mice 4-5 weeks of age were maintained on a standard mouse diet. Treatment groups of 5 mice each were as follows: a group treated with ISIS 394814; a control group treated with saline; and control group treated with an oligonucleotide not complementary to any known gene sequence (ISIS 141923). Oligonucleotide or saline was administered intraperitoneally twice weekly, for a period of 6 weeks; oligonucleotide doses were 50 mg/kg (100 mg/kg/wk total). Real-time PCR of liver mRNA and serum analyses were performed as described for the C57Bl/6 study.

Antisense inhibition of PCSK9 reduced liver PCSK9 mRNA by approximately 90%, relative to saline controls. However, no reduction in serum cholesterol was observed. Liver triglyceride levels were unaffected by antisense inhibition of PCKS9 in these mice. An 80% increase in liver apobec1 mRNA levels was observed, relative to saline controls. These data suggest that a functioning LDL-R is required for cholesterol and liver triglyceride reduction via PCSK9 antisense inhibition.

Example 13 Antisense Reduction of PCSK9 mRNA in Mouse Primary Hepatocytes

Antisense oligonucleotides targeted to PCSK9 were tested for antisense inhibition of PCSK9 in mouse primary hepatocytes. Mouse primary hepatocytes were purchased from a commercial supplier, and cultured according to routine culture procedures. Cells were treated with 6.25, 12.5, 25, 50, 100, or 200 nM of antisense oligonucleotide for a period of 24 hours, after which RNA was isolated and PCSK9 mRNA was measured by real-time PCR as described herein.

The antisense oligonucleotides tested in human primary hepatocytes were Isis 395165, 395185, 395186, 395187, 405879, 405881, 405891, 405988, 405994, and 406008. The antisense oligonucleotides inhibited PCSK9 in a dose-dependent manner in human primary hepatocytes as shown in Table 26.

TABLE 26 Dose-Dependent Reduction PCSK9 mRNA in Mouse Primary Hepatocytes 6.25 200.0 ISIS No nM 12.5 nM 25.0 nM 50.0 nM 100.0 nM nM 395165 95.2 94.9 81.7 66.3 57.5 29.9 395185 109.2 100 90.5 81.2 64.7 35.8 395186 101.9 91.5 77.8 59.1 39.5 13.7 395187 102.4 96.1 83.9 66.4 40 15.3 405879 107.3 97.5 87.8 76.6 63 43.1 405881 104.8 102.1 98.1 84.9 68.7 48.9 405891 104.9 103.3 101.5 88.9 79.6 49.5 405988 97.3 101.1 91.7 82.3 61.2 34.8 405994 110.2 101 111.6 94.4 80.1 57.7 406008 99.5 91.3 83.9 73.7 61.8 39.4 157700 97.8 98.3 96.2 94.3 83.2 76.4 141923 84.3 80 79.9 77.3 98.8 64.2

Example 14 Antisense Reduction of Human PCSK9 mRNA in Hep3B and HeLa Cells: Dose Response Experiment

A subset of the antisense oligonucleotides that inhibited PCSK9 expression at high levels in the cell culture systems described above was selected for further screening. The group of selected antisense compounds is shown in Table 27. Table 27 shows the Isis number, nucleotide sequence, and percent inhibition obtained in Hep3B cells using the protocol disclosed in Example 3 (see Table 17) for the various antisense compounds.

TABLE 27 SEQ % ID Isis No. Sequence 5′-3′ Motif inhibition NO 405881 CACCCTTGGCCACGCCGGCA 5-10-5 98.3 250 399819 CCCACTCAAGGGCCAGGCCA 5-10-5 93.4 65 395165 ACCCTTGGCCACGCCGGCAT 5-10-5 96.7 28 405879 CCTTGGCCACGCCGGCATCC 5-10-5 94.1 248 406008 CTTGGTGAGGTATCCCCGGC 5-10-5 95.2 188 405891 TCCTCAGGGAACCAGGCCTC 5-10-5 96.1 352 395186 CTTTGCATTCCAGACCTGGG 5-10-5 93.7 60 405988 GGCAGCACCTGGCAATGGCG 5-10-5 95.9 381 405994 GCAGTGGACACGGGTCCCCA 5-10-5 95.2 400 406023 TGGTATTCATCCGCCCGGTA 5-10-5 92.5 212 395187 GGCAGCAGATGGCAACGGCT 5-10-5 92.7 62 395185 CACGGGTCCCCATGCTGGCC 5-10-5 92.7 59 406033 CCTGCCAGGTGGGTGCCATG 5-10-5 91.2 237 405923 GGCATTGGTGGCCCCAACTG 5-10-5 94.3 288 399900 GGTCCTCAGGGAACCAGGCC 3-14-3 92.0 50 405995 TGGCAGTGGACACGGGTCCC 5-10-5 92.4 402 405991 GACACGGGTCCCCATGCTGG 5-10-5 91.9 394 406005 CGGGCAGTGCGCTCTGACTG 5-10-5 83.4 180 399793 CCTCGGAACGCAAGGCTAGC 5-10-5 77.9 8 395152 ACGCAAGGCTAGCACCAGCT 5-10-5 93.3 7

The IC₅₀ of the various antisense compounds was determined using different cell culture systems. Antisense oligonucleotides targeted to PCSK9 were tested at various doses in Hep3B or HeLa cells, for example. Cells were plated at densities of 4,500 cells or 5,000 per well, respectively, and treated with of antisense oligonucleotides at various concentrations. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein, relative to untreated control cells. Two different human PCSK9 primer probe sets were used to measure mRNA levels. Results with Primer Probe Set 2740 are shown in Table 28. PCSK9 mRNA levels were adjusted according to total RNA content as measured by RIBOGREEN®. Table 28 shows IC₅₀ values (nM) obtained from two experiments with Hep3B cells and from one experiment with HeLa cells.

TABLE 28 IC₅₀ values Hep3B Hep3B HeLa SEQ IC₅₀ IC₅₀ IC₅₀ ID ISIS # (nM) (nM) (nM) PCSK9 Target Sequence NO 405881 21 26 13 CACCCTTGGCCACGCCGGCA 250 399819 21 28 12 CCCACTCAAGGGCCAGGCCA 65 395165 23 28 13 ACCCTTGGCCACGCCGGCAT 28 405879 24 28 14 CCTTGGCCACGCCGGCATCC 248 406008 31 35 8 CTTGGTGAGGTATCCCCGGC 188 405891 32 38 14 TCCTCAGGGAACCAGGCCTC 352 395186 33 33 7 CTTTGCATTCCAGACCTGGG 60 405988 34 42 8 GGCAGCACCTGGCAATGGCG 381 405994 35 39 9 GCAGTGGACACGGGTCCCCA 400 406023 40 50 18 TGGTATTCATCCGCCCGGTA 212 395187 41 44 8 GGCAGCAGATGGCAACGGCT 62 395185 42 43 11 CACGGGTCCCCATGCTGGCC 59 406033 44 61 15 CCTGCCAGGTGGGTGCCATG 237 405923 47 41 20 GGCATTGGTGGCCCCAACTG 288 399900 48 44 21 GGTCCTCAGGGAACCAGGCC 50 405995 48 63 12 TGGCAGTGGACACGGGTCCC 402 405991 63 52 15 GACACGGGTCCCCATGCTGG 394 406005 63 CGGGCAGTGCGCTCTGACTG 180 399793 >75 CCTCGGAACGCAAGGCTAGC 8 395152 46 16 ACGCAAGGCTAGCACCAGCT 7

Several of the antisense compounds examined above were selected for further study. These antisense compounds exhibited at least about 93% inhibition of human PCSK9 mRNA expression in the Hep3B cell culture system, as shown in Tables 17-19, for example. As shown in Table 28, the selected antisense compounds also displayed IC₅₀ parameters of less than about 45 nM when tested with Hep3B cells and less than about 15 nM when tested with HeLa cells. The selected antisense compounds, listed in Table 29, are Isis 405881, Isis 395165, Isis 405879, Isis 406008, Isis 405891, Isis 395186, Isis 405988, Isis 405994, Isis 395187, and Isis 395185. Isis 405879, for example, showed superior results in both Hep3B and HeLa cells, with IC₅₀ values of about 26 nM and 14 nM, respectively.

Example 15 Dose-Dependent Antisense Reduction of PCSK9 mRNA in Human Primary Hepatocytes; Cynomolgus Monkey (“Cyno”) Primary Hepatocytes; and Human HEK-293 Cells

The selected antisense oligonucleotides were further tested for dose dependent antisense inhibition in human primary hepatocytes, cyno primary hepatocytes, and HEK-293 cells expressing human PCSK9. Cells were purchased from commercial suppliers and cultured according to routine culture procedures. Cells were treated with 10, 25, 50, 150, or 300 nM antisense oligonucleotide (i.e., a five-point dose response test) for a period of 24 hours, after which RNA was isolated. PCSK9 mRNA was measured by real-time PCR, as described herein.

Results of the five-point dose response tests using HEK-293 cells are shown in FIG. 2. Results of the five-point dose response tests using cyno primary hepatocytes are shown in FIG. 3. The IC₅₀ results, expressed in mM units, for the five-point dose response test using human primary hepatocytes, cyno primary hepatocytes, HEK-293 cells, Hep3B cells, and HeLa cells are summarized in Table 29.

TABLE 29 IC₅₀ (nM) for Selected Antisense Compounds Isis No. 395165 395185 395186 395187 405879 405881 405891 405988 405994 406008 SEQ ID NO: 28 59 60 62 248 250 352 381 400 188 Human Primary 24 33 39 30 ND 50 297 166 56 163 Hepatocytes Cyno Primary 5 17 18 14 2 19 10 12 21 11 Hepatocytes HEK-293 40 116 6 11 35 86 21 11 17 27 Hep3B 28 43 33 44 28 26 38 42 39 35 HeLa 13 11 7 8 14 13 14 8 9 8

ISIS 405879 was tested at various doses in human primary hepatocytes. RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. Human PCSK9 primer probe set 2740 was used to measure mRNA levels. Results are shown in Table 29.1.

TABLE 29.1 Dose-dependent Reduction in Human PCSK9 mRNA Expression in Human Primary Hepatocytes 300 200 ISIS NO. nM nM 150 nM 100 nM 50 nM 25 nM 10 nM 405879 13 18 26 30 27 54 75

Isis 405879 has an IC₅₀ value of 29 nM in human primary hepatocytes. Overall Isis 405879 displays superior activity, particularly in cyno primary hepatocytes. Isis 405879 was thus further tested in the various cell models, and the results of the additional testing are shown in Table 30. In these experiments, Isis 405879 was administered to the cells using lipofectin transfection reagents. PCSK9 mRNA was assayed by PCR following 24 hr incubation after transfection. These results confirm that Isis 405879 potently inhibits PCSK9 mRNA expression at low concentrations.

TABLE 30 Isis 405879 activity in cell culture models Hepatocyte Transgenic Hep3B HepG2 HeLa Transgenic Primary Primary HEK293 Cells Cells Cells Mouse Hepatocyte Hepatocyte cells (human) (human) (human) (h-PCSK9) (human) (cyno) (h-PCSK9) Mean 17 5 14 140 22 7.4 18 IC₅₀ (nM)

These experiments confirm that the presently disclosed antisense oligonucleotides are capable of potently inhibiting PCSK9 mRNA expression in a variety of cell model systems, including a human primary cell line. Exemplary antisense compounds inhibit PCSK9 mRNA expression with IC₅₀ values in the nM range.

The transgenic PCSK9 cDNA construct expressed in mouse hepatocytes is thought to be rapidly exported to the cytoplasm, because it does not require post-transcriptional editing. The antisense oligonucleotides provided herein act by recruiting the RNase H enzyme, which is restricted to the nucleus. Therefore, inhibition by antisense oligonucleotides in this cell type is significantly diminished.

Example 16 In Vivo Testing Procedures 16.1. High Fat (HF)-Fed Hyperlipidemic Mouse Model

Antisense compounds may be tested in a high fat-fed, hyperlipidemic mouse model to test efficacy of the antisense compounds in reducing PCSK9 mRNA expression, increasing LDL-R expression, reducing total cholesterol and LDL, and affecting other metrics of antisense compound efficacy. A representative hyperlipidemic mouse model is described in Graham et al., J. Lipid Res. (2007) 48: 763-767.

All animal experiments were conducted in accordance with Institutional American Association for the Accreditation of Laboratory Animal Care guidelines. C57BL/6 mice were obtained from Jackson Laboratory (http://www.jax.org). A majority of the mice were male, and studies were initiated when animals were 4-5 weeks of age. The mice were maintained on a 12 h light/12 h dark cycle and fed ad libitum. C57BL/6 mice were fed a diet consisting of 60% lard (Research Diets, New Brunswick, N.J.), whereas experimental Ldlr-deficient/apoB-100 mice were fed regular chow.

Antisense compounds were administered twice weekly (50 mg/kg) for 6 weeks by intraperitoneal injection (10 mg/mL dosing solution formulated in saline; saline control). After the treatment period, whole blood was collected for analysis of serum parameters, and whole liver was collected for RNA analysis, protein analysis, and histological evaluation.

16.2. H-ApoB/CETP Transgenic Mouse Model

A transgenic mouse model that expresses both human apolipoprotein (apo) B and human cholesteryl ester transfer protein (CETP) provides another useful animal model system to test the efficacy of the presently disclosed antisense compounds. The lipoprotein cholesterol distribution in the serum of a chow-fed h-apoB/CETP mouse resembles a human profile. Specifically, the percentages of total cholesterol within the HDL, LDL, and VLDL fractions of apoB/CETP animals are approximately 30%, 65%, and 5%, respectively, similar to the distribution of cholesterol in the plasma of normolipidemic humans. The h-apoB/CETP mouse model is described further in Grass et al., J. Lipid Res. (1995) 36: 1082-1091.

16.3. H-PCSK9 Transgenic Mouse Model

A transgenic mouse expressing human PCSK9 provides another useful animal model to study in vivo repression of PCSK9 mRNA by the present antisense compounds. H-PCSK9 transgenic mice overexpress human PCSK9 in liver and secrete large amounts of the protein into plasma, which increases plasma LDL-C concentrations to levels similar to those of LDLR-knockout mice. H-PCSK9 transgenic mice were constructed by inserting a cDNA encoding human PCSK9 into a pLiv-11 vector that contained the constitutive human apoE promoter and its hepatic control region, as described in Simonet et al., J. Biol. Chem. (1993) 268: 8221-8229. Transgenic mice were generated by injecting linearized pLiv-11-hPCSK9 into the fertilized eggs as described in Shimano et al., J. Clin. Invest. (1996) 98: 1575-1584. H-PCSK9 transgenic mice are further described in Lagace et al., J. Clin. Invest. (2006) 116: 2995-3005.

16.4. Mouse and Rat Models for In Vivo Tolerability, Half-Life and Tissue Distribution

Mouse (CD1) and rat (Sprague-Dawley) models were used to test the in vivo tolerability of various antisense compounds. In the mouse CD1 model, antisense compounds were overdosed at 100 mg/kg/wk for six weeks. Possible proinflammatory effects and hepatotoxicity were assayed following administration of the antisense compounds. Serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN), creatinine, albumin, and bilirubin were assayed. Weights of livers, kidneys, and spleens of animals were determined. Histopathologic screening was conducted to detect hepatocellular swelling, multifocal apoptosis, inflammatory cell infiltrates in the liver or lungs, or follicular hyperplasia. Additional studies were conducted in the rat model to screen for renal toxicity or high renal accumulation and proinflammatory effects. In these experiments, antisense compounds were administered at 60 or 100 mg/kg/wk for six weeks. In addition to the markers examined above, the urine protein/creatine ratio and proteinuria were assayed.

The half-life of administered antisense compounds also was examined in the target organ, i.e., liver, in the mouse model. For these experiments, antisense compounds were administered twice a week for two weeks at 50 mg/kg, and target organs were then harvested and assayed for the presence of the antisense compounds, as described below.

The distribution of administered antisense compounds in various organs also was examined in the rat model. Rats in these studies were administered 20 mg/kg antisense compounds twice weekly for three weeks. Livers and kidneys of test animals were then harvested and assayed for the presence of the antisense compounds, as described below.

16.5. Cynomolgus Monkey (M. fascicularis) Model

Potencies and pharmacodynamics of antisense compounds were compared in a cyno model to identify the compounds exhibiting the best combination of high potency and low renal accumulation, when given subcutaneously to animals for a 5-week period. In another study, dosing was conducted over a 13-week period. The cyno monkey is widely used in toxicity testing, and a substantial historic toxicology database is available for this model.

The PCSK9 target sequence for an antisense compound may differ in humans versus cyno monkey. As a result, an antisense compound for use in humans may not be fully complementary to a monkey PCSK9 nucleic acid. Antisense compounds having identical target sequences in the two species are preferred. Isis 405881 and Isis 395165 each have a one nucleotide mismatch compared to the cyno sequence, meaning that these antisense compounds are less preferred in the cyno model.

Cyno monkeys (Charles River BRF, Inc.) were approximately 26 to 32 months old and 2.5 to 4 kg in weight on the first day of dosing. Monkeys were offered water ad libitum and fed a daily ration of 12 biscuits approximately 2 hours post dose. Antisense compounds were administered subcutaneously at 3 doses every other day during the first week, then by weekly injections. A low dose of 15 mg/kg was used to assess the potency of the antisense compounds. A high dose of 8 subcutaneous injections of 30 mg/kg over 5 weeks or 13 weeks was used to assay renal accumulation and pharmacodynamics. Antisense compounds were administered at a concentration of 60 or 120 mg/mL, respectively, in a vehicle of sterile phosphate buffered saline. Plasma concentrations of antisense compounds were determined at each dose level to estimate systemic exposure in monkeys following subcutaneous administration and to assess whether systemic exposure was altered after repeat dosing.

To measure the host reaction to the administered antisense compounds, 2 mL blood samples were collected from the femoral vein of host individuals in tubes of appropriate size containing K₂EDTA. Plasma was obtained by centrifugation and samples were stored frozen. The analysis of the plasma samples included a determination of the level of plasma PCSK9 protein. To measure an effect on pharmacodynamic end points, LDL, HDL, and apoB levels also were analyzed. To determine whether the host individuals suffered side effects, blood samples were further analyzed for an effect on the hematological parameters listed in Table 31 and the serum chemistry parameters listed in Table 32:

TABLE 31 Hematology markers erythrocyte count red cell distribution width hemoglobin platelet count hematocrit mean platelet volume mean corpuscular volume absolute total and differential leukocyte counts mean corpuscular hemoglobin evaluation of cell morphology concentration absolute reticulocyte count

TABLE 32 Serum chemistry markers aspartate aminotransferase triglycerides alanine aminotransferase glucose gamma glutamyltransferase urea nitrogen alkaline phosphatase creatinine total bilirubin calcium total cholesterol phosphorus total protein sodium albumin potassium globulins chloride albumin/globulin ratio

Possible additional side effects were determined by screening individuals administered antisense compounds for gross changes in organs or microscopic changes in tissues. A limited necropsy conducted on all test animals included gross examination of various organs and tissues. Terminal body weights were recorded, and kidneys and spleen were weighed for all test animals. For microscopic evaluation, liver, kidney, spleen and injection sites from all animals were fixed in 10% neutral buffered formalin and were processed, embedded in paraffin, sectioned, and stained with hematoxylin and eosin.

Representative tissue samples of liver and kidney cortex from each scheduled-necropsy monkey from the high-dose and control groups were collected for ultrastructural examination using electron microscopy. Thin slices (2-4 mm in thickness) of liver and kidney cortex were cut and transferred to a piece of dental wax containing a small volume of McDowell-Trump fixative. Thin slices were further minced into approximately 1×1-2 mm cubes in McDowell-Trump fixative and analyzed by electron microscopy.

To evaluate PCSK9 mRNA levels, samples from liver (approximately 1 g) were homogenized using a Polytron tissue disrupter in 10 mL of RLT solution (Qiagen) and snap-frozen and stored at −70° C. or colder. Additionally, samples (approximately 500 mg each) from kidney were collected and snap-frozen for mRNA analysis.

Tissue samples from liver and kidney cortex were also analyzed for the presence of the administered antisense compounds. Antisense compounds present in the tissues can be detected and quantified using capillary gel electrophoresis (CGE), as described in Leeds et al, Drug Metab Dispos. (1998) 26: 670-675, for example. In this procedure, tissues are weighed, mixed with an internal standard (e.g., a 27-mer (T) oligonucleotide) in proteinase K digestion buffer (2.0 mg/ml proteinase K in 20 mM Tris-HCl, pH 8.0, 20 mM EDTA, 100 mM NaCl, 0.5% Nonidet P-40), and homogenized. Overnight incubation at 37° C. is followed by phenol/chloroform extraction. Tissue extracts are purified by sequential anion-exchange solid-phase extraction (SPE) followed by reverse-phase SPE, described in Leeds et al., Anal Biochem (1996) 235: 36-43. Samples are further desalted by membrane dialysis before analysis using gel-filled capillaries, as described in Leeds et al. (1996).

Capillary gel electrophoresis separations can be performed as described by Leeds et al. (1998), using a Beckman P/ACE capillary electrophoresis instrument (model 5010) with a 27-cm column (effective length, 20 cm) containing 12% polyacrylamide, with 8.3 M urea in 100 mM Tris-borate, pH 8.5, as the running buffer. Separation is achieved at 50° C. and 550 V/cm. Oligomers eluting from the column are detected by UV absorption at 260 nm. Quantification of antisense oligonucleotides is based on the tissue weight extracted and the initial T27 concentration.

Example 17 In Vivo Testing Results 17.1. Reducing PCSK9 mRNA In Vivo Using a Murine Antisense Compound Correlates with In Vivo Plasma LDL-C and Liver TG Reduction

In vivo pharmacodynamic studies were performed with the murine antisense compound Isis 394816 to validate the pharmacodynamic endpoint of PCSK9 mRNA reduction. Isis 394816 has the sequence 5′ GGTAAGGTGCGGTAAGTCCT 3′ (SEQ ID NO: 462), which targets position 3100 (5′) of the coding sequence of murine PCSK9 mRNA (NCBI Accession No. NM_(—)153565.1; SEQ ID NO: 463). Isis 394816 has a 3-14-3 motif.

Isis 394816 was administered by intraperitoneal injection to an h-apoB/CETP transgenic mouse for 6 weeks at a dosing regimen of 20 mg/kg/wk or 50 mg/kg/wk. See Example 16.2. As shown in FIG. 4, Isis 394816 administered at either dose caused an approximate 80% reduction in PCSK9 mRNA expression in livers of test animals, compared to control animals. As shown in FIG. 5, panel A, Isis 394816 produced a significant (p<0.0001) decrease in the level of plasma LDL-C at the end of six weeks. Additionally, as shown in FIG. 5, panel B, Isis 394816 produced a significant (p<0.02) decrease in the level of liver TG at the end of six weeks.

Additionally, Isis 394816 reduced PCSK9 mRNA expression, plasma LDL-C, and liver TG in a dose dependent manner, as shown in FIG. 6. The potency of Isis 394816 was much higher in a hyperlipidemic mouse model (see Example 16.1.) than in a lean mouse model (data not shown). The reduction of plasma LDL-C positively correlated with the inhibition of PCSK9 mRNA levels in liver, shown in FIG. 7. These studies demonstrate that the reduction of liver PCSK9 mRNA correlates with the desired endpoint of reducing plasma LDL-C and liver TG. Accordingly, the reduction of liver PCSK9 mRNA provides a valid pharmacodynamic endpoint.

17.2. Differences in the Half-Life (t_(1/2)) of Antisense Compounds in Liver

The half-lives of selected antisense compounds were assayed in mouse CD1 livers, according to the procedures set forth in Example 16.4. Half-lives varied among the tested antisense compounds, which again is attributable to differences in structural features of the antisense compounds, such as nucleotide sequences and/or motifs. For example, the half-life of Isis 405879 (t_(1/2)=27.7 days) was significantly higher than the half-life of Isis 395165 (20.4 days), although the target sequences of these two antisense compounds are shifted by only two nucleotides:

Isis 405879: CCTTGGCCACGCCGGCATCC Isis 395165: ACCCTTGGCCACGCCGGCAT Half-life data for exemplary antisense compounds is summarized in Table 33. Isis 405879 displayed a particularly high half-life in liver.

TABLE 33 Liver t_(1/2) (days) Antisense Compound Liver t_(1/2) (d) 395165 20.4 395185 20.2 395186 19.9 395187 13.7 405879 27.7 405881 14.1 405891 17.1 405988 14.6 405994 20.2 406008 19.7

17.3. Differences in the Liver and Kidney Distributions of Antisense Compounds

The antisense compounds listed in Table 29 were administered to rats to determine the distribution of antisense compounds in liver and kidney. Accumulation of high levels of antisense compounds in the kidneys may cause adverse side effects. Antisense compounds were dosed and administered as described in Example 16.2. FIG. 8 shows the distribution of the antisense compounds shown in Table 29 in rat liver and kidneys. Isis 395185, Isis 395186, Isis 395187, Isis 405879, Isis 405881, Isis 405891, Isis 405988, and Isis 405994 show a particularly advantageous ratio of distribution between liver and kidney.

When tissue distribution was tested in a cyno model, further differences among antisense compounds were observed. Accumulation of Isis 395186, Isis 405879, and Isis 405891 in liver and kidney was examined after dosing each antisense compound at two concentrations. See Example 16.5. The results are shown in FIG. 9. Accumulation at each dosing concentration (15 mg/kg and 30 mg/kg) is shown for kidney (left side bar) and liver (right side bar). Isis 405879 advantageously accumulates at a high concentration in liver, even at a dose of 15 mg/kg, and accumulates at a low level in kidney. Specifically, Isis 405879 accumulated in kidneys at a concentration of 1300 μg/g (15 mg/kg dose) to 1800 μg/g (30 mg/kg dose). The average accumulation of antisense compounds in kidneys is 2500 μg/g (15 mg/kg dose) to 4000 μg/g (30 mg/kg dose). Variation in kidney and liver accumulation is attributable to structural differences, such as nucleotide sequences and/or motifs, between the antisense compounds.

17.4. Differences in Host Tolerance of Antisense Compounds

Rodent tolerability studies, conducted as described in Example 16.4, showed that antisense compounds were well tolerated. Following administration of the exemplary antisense compounds listing in Table 29, serum chemistry markers were normal, with some variably high ALT and AST levels observed for Isis 405988 and Isis 405994. Whole body weight and major organ weights in CD-1 mice also were not significantly affected by administration of the exemplary antisense compounds. Histological findings revealed differences between the various antisense compounds, but were consistent with the overall good tolerability for the antisense compounds. The results of the histological screens are shown in Table 34.

17.5. Results of the 5 Wk Cyno Toxicology Study

The methods used to conduct the 5 wk cyno toxicology study are described in Example 16.5. Isis 405879 is referred to as BMS-844421 below and in the corresponding figures. The PCSK9 mRNA levels assayed by q-RT-PCR in the cyno liver tissue samples were decreased by BMS-844421 treatment by an average of 65% and 70% (average of males and females) versus saline controls at the low and high doses, respectively (FIG. 25). The effect was seen in both females and male monkeys, despite relatively high variability of mRNA level between animals. No dose-dependency was seen between the 2 doses. The data are mean+/− for male and female animals combined.

The levels of BMS-844421 ASO were measured in liver at the end of the 5 wk study. As shown in FIG. 26, liver full-length BMS-844421 levels reached 1 mg/gram tissue in both dose groups. This level is consistent with values from previous experiences with other liver-targeting ASOs. Most of the compound was detected as intact full-length BMS-844421. No dose-dependency was seen in the ASO liver accumulation data, in agreement with the lack of dose-dependent effects on liver PCSK9 target mRNA.

Pharmacodynamic responses were assessed in serum lipid assays for the cyno 5 wk ASO study. As seen in FIG. 27, a modest trend (not statistically significant) toward lowering LDL-C was observed for BMS-844421 treated cyno monkeys at 2.5 and 5 wk of dosing. This trend at both time points for BMS-844421 relative to baseline was also seen in the scatterplot of individual animal data in FIG. 28A. HDL/LDL ratios were sometimes employed as a sensitive indicator of plasma lipid status. A chart of the average ratio of HDL/LDL in the cyno study showed a trend for increase in the ratio for BMS-844421 treated cynos at 2.5 and 5 wk (FIG. 28B).

Gene expression profiling by Affymetrix microarray was conducted on the RNA samples from this study. The analysis included liver RNA from animals administered vehicle alone, BMS-844421 at 15 mg/kg and 30 mg/kg and BMS-844419 and BMS-844423 at 30 mg/kg only. By microarray, there were significant (p<0.01) transcriptional changes in approximately 1 to 2% of genes with approximately 1% expected to change at this significance level by chance. For drug-dosed groups relative to control there were alterations in inflammation-, immune system-, and lipid homeostasis-related transcripts with statistically significant animal to animal variability. Other drug-related hepatic transcriptional changes common to multiple dose groups were associated with several or a few genes in various biological processes and are of unknown significance.

17.6. Results of the 13 Wk Cyno Toxicology Study

An additional opportunity to monitor pharmacodynamic responses to BMS-844421 in monkeys was the 13 wk toxicology study, described in Example 16.5. Lean cyno monkeys with low LDL levels were again the subject for this study, as for the 5 wk cyno monkey study above. At 13 wks subcutaneous dosing at 4 different dose levels, BMS-844421 suppressed PCSK9 expression as measured by both liver PCSK9 mRNA and plasma PCSK9 protein levels. Despite the inter-animal variability in these values, it is apparent that BMS-844421 strongly suppressed both liver mRNA and plasma PCSK9 protein (FIG. 29). 40 mg/kg suppressed PCSK9 protein levels very robustly with less variation between animals (FIG. 29). No statistically significant differences were seen in plasma LDL-C or other lipid parameters in this study.

TABLE 34 Histological Findings in CD-1 Mice Antisense Lymph Skeletal compound Liver Kidney Lung Heart Spleen node Intestine Thyroid Sternum Muscle Saline − − − − − − − − − − ISIS 395165 +/++ − −/+ −/+ − − − − ISIS 395185 + − − − −/+ − − − − − ISIS 395186 − − − − − − − − − − ISIS 395187 − − − − −/+ − − − − − ISIS 405879 − − − − − − − − − − ISIS 405881 inflam − inflam − −/+ −/+ − − − − ISIS 405891 − − − − −/+ −/+ − − − − ISIS 405988 − − − − − − − − − − ISIS 405994 ++ − − − −/+ − − − − − ISIS 406008 +/++ − − − − − − − − − Normal: − Minimal changes: −/+ Mild changes: + Moderate changes: ++ Severe changes: +++

Microscopic evaluation of mouse organs from animals treated with 10 different human PCSK-9 antisense compounds revealed that all compounds generally were well tolerated, but that some antisense compounds were particularly well tolerated. The following results were seen in the various examined organs.

-   -   Liver: Mild (+) to moderate (++) liver injuries (cytoplasmic         swelling to multifocal apoptosis) were seen in Isis 395165         (+/++), Isis 395185 (+), Isis 405988 (+/++), Isis 405994 (++),         and Isis 406008 (+/++) treated livers. Mild inflammatory changes         were seen in Isis 405881 treated livers.     -   Lung: No significant abnormality was seen in treated animals         except for mild histiocytosis noticed in Isis 405881 treated         animals.     -   Spleen: Mild follicular hyperplasia was seen in Isis 395165,         Isis 395185, Isis 395187, Isis 405881, Isis 405891, and Isis         405994 treated spleens.     -   Lymph node: Minimal histiocytosis was observed in Isis 395165,         Isis 405881 and Isis 405891 treated animals.     -   Heart: Minimal levels of macrophage infiltration were seen in         Isis 395165, Isis 395186, Isis 405879, Isis 405881, Isis 405891,         Isis 405988, and Isis 406008 treated hearts.     -   Kidney, Thyroid, small intestine, skeletal muscle and sternum:         No abnormality was visualized.

Differences in the histological findings were seen even between Isis 395165 and Isis 405879. As noted above, the target sequences of these two antisense compounds are only shifted by two nucleotide positions. Nevertheless, Isis 405879 advantageously displays no adverse reactions, whereas Isis 395165 elicits a mild inflammatory response in liver, spleen, and lymph nodes. In summary, Isis 405879, Isis 395186, and Isis 405988 display superior tolerance, compared to the other antisense compounds tested in this study.

17.7. Differences in Achieving Pharmacodynamic Endpoints in the Cyno Model

Isis 395186 (“844419”), Isis 405879 (“844421”), and Isis 405891 (“844423”) were selected for further study in the cyno model, using the procedures described in Example 16.6. As set forth above, these antisense compounds displayed high activity in cell culture models, were well tolerated, and displayed advantageous tissue distribution and half-life properties. Further, none of these antisense compounds targeted a PCSK9 sequence known to have differences in monkeys and humans. In addition, the target sequences in human PCSK9 were known not to contain SNPs having a wide distribution in the population.

The antisense compounds were tested for their ability to reduce PCSK9 mRNA in cyno liver. See Example 16.6. The results of this experiment are shown in FIG. 10. Despite variability between the tested animals, Isis 405879 (labeled “844421”) displayed a superior ability to reduce PCSK9 mRNA levels significantly. When administered at a low dose (15 mg/kg, shown in the left side bar), Isis 405879 significantly (p<0.023) reduced PCSK9 mRNA levels. When administered at a high dose (30 mg/kg, shown in the right side bar), Isis 405879 also significantly (p<0.011) inhibited PCSK9 mRNA expression. The results obtained with Isis 405879 were superior to those obtained with Isis 395186 (labeled “844419”) and Isis 405891 (labeled “844423”). As shown in FIG. 11, Northern analysis of liver PCSK9 mRNA confirms that 15 mg/kg and 30 mg/kg Isis 405879 substantially reduces PCSK9 mRNA levels.

As described in Example 17.1, the reduction of PCSK9 mRNA is expected to correlate with a reduction in plasma LDL-C and liver TG. Consistent with this expectation, Isis 405879 produced a significant (p<0.06) decrease in cyno serum LDL-C after five weeks of administration at 30 mg/kg, as shown in FIG. 12. Isis 405879 additionally produced a significant (p<0.05) increase in the ratio of HDL to LDL following five weeks of administration at 30 mg/kg, as shown in FIG. 13.

Generally confirming earlier data obtained in rodents, Isis 405879 elicited no differences in serum chemistry, hematology, or organ weights, except for a slight increase in spleen weight. Histology revealed indicia of only mild inflammation. The advantageous distribution of Isis 405879 in liver and kidney in the cyno model is described above and shown in FIG. 9.

Example 18 Dose Response Study of Isis 405879 (“844421”) in a Human PCSK9 Transgenic Mouse Model

BMS-844421 is a human specific PCSK9 antisense oligonucleotide (ASO) designed to downregulate hepatic PCSK9 expression, increase LDL receptor activity, and promote clearance of LDL and reduction of plasma LDL-C levels in humans. A human genomic PCSK9 transgenic mouse model was recently established. This mouse line expresses hemizygous transgene in the presence of endogenous mouse PCSK9 (i.e., both genes are simultaneously expressed). It was found that the transgenic PCSK9 mice responded to chronic dosing with BMS-844421 and related human PCSK9 ASOs with decreased liver mRNA expression and decreased circulating human PCSK9 protein levels.

The present study examined the dose response to BMS-844421 in the hemizygous genomic transgenic mouse model on a normal chow diet. Experiments were conducted comparing the effects of BMS-844421 (human specific PCSK9 ASO) to those of ISIS 394816 (murine specific PCSK9 ASO) in this model alone and in combination. BMS-844421 exhibited dose-dependent suppression of liver target mRNA, reflected in dose-dependent decreases in plasma human PCSK9 levels. Both liver mRNA and plasma protein gave ED50 values of approximately 15 mg/kg (given twice weekly) for BMS-844421, while the suppression of plasma PCSK9 protein was greater than liver PCSK9 mRNA at each dose in this model. Trends for increased LDLR protein in liver total membrane fraction, and a feedback effect on liver HMGCoA reductase mRNA expression were observed, but no changes in plasma lipids were seen for BMS-844421 treated hemizygous mice. The combination of ISIS 394816+BMS-844421 (each at 15 mg/kg) elicted a greater increase in liver LDLR protein than either ASO alone, consistent with the proposed mechanism of action and the dual expression of both human and murine PCSK9 in this model.

18.1. Materials and Methods

The sequences of BMS-844421 and other ASOs tested in this report are given in Table 35. The negative control ISIS 141923 is not complementary to any known gene. The design of the dose response study is described in the beginning of the Results section (section 18.2. below).

TABLE 35 Nucleotide sequences of ASOs ASO Sequence BMS 844421, human PCSK9 5′-CCTTGGCCACGCCGGCATCC-3′ specific* ISIS 141923, 5′-CCTTCCCTGAAGGTTCCTCC-3′ non-targeting ISIS 394816, murine 5′-GGGCTCATAGCACATTATCC-3′ PCSK9 specific *BMS-844421 is identical to ISIS 405879

Parameters evaluated at study termination included hepatic mRNA, hepatic LDLR protein levels, plasma concentrations of total cholesterol, LDL, HDL, triglycerides, transaminases, and organ and body weights. Whole liver was processed for RNA, and protein, and held for possible histological examination and hepatic triglyceride levels. Blood and liver samples were obtained 24 hr after the prior dose. Blood samples were obtained from tail vein or retro orbital plexus and EDTA, chilled on ice, and plasma samples were obtained by standard procedures. For mRNA assays, liver samples (30-50 mg each) were placed in eppendorf tubes containing 1000 uL of TRIzol (Invitrogen #15596-018) and homogenized using stainless steel beads on a Tissue Lyser instrument (Qiagen, Valencia, Calif.). Chloroform (200 μL) was added to each tube, the contents were mixed vigorously by hand and tubes were centrifuged at 12,000×g for 20 minutes at 4° C. About 300 μL of each supernatant was removed and placed in 1.5 mL microcentrifuge tubes. One volume of 70% EtOH was added, the mixtures were vortexed thoroughly and transferred to RNeasy Mini Spin Columns (QIAgen Cat. No. 74106). RNA was purified, treated with DNase and eluted into microcentrifuge tubes following QIAgen's manufacturer's instructions.

Plasma human PCSK9 levels were assayed using the human specific ELISA with mAb 4H5 as capture antibody.

18.2. Results

A dose-response study was run in human PCSK9 genomic transgenic mouse line 66 mice on a normal chow diet. Groups of 6 mice (3 males, 3 females) received the following treatments for 6 weeks. BMS-844421 was administered p.o. at 5, 10, 15, and 30 mg/kg per dose given twice weekly (on Tuesdays and Fridays) for 6 weeks. Controls received saline, and an additional group received the non-targeting negative control ASO ISIS 141923 (30 mg/kg twice weekly). The murine PCSK9 ASO ISIS 394816 was administered to another group at 15 mg/kg per dose. The final group received BMS-844421 (15 mg/kg) plus ISIS 394816 (15 mg/kg) combined.

Liver mRNA assayed by qRT-PCR showed that BMS-844421 decreased liver PCSK9 mRNA in a dose dependent manner, with greatest effect at the top dose studied (FIG. 14). The negative control ASO, ISIS 141923, had no significant effect on target mRNA, while the murine targeted ASO, ISIS 394816, decreased human PCSK9 mRNA by 33%, suggesting partial cross-hybridization to human PCSK9 mRNA.

The ASO effects on plasma human PCSK9 followed a similar pattern as mRNA, while the effects of the higher doses of BMS-844421 resulted in greater effects, up to 70% suppression at 30 mg/kg/dose (FIG. 15). ISIS 394816 suppressed approximately 30% (similar to mRNA effects), while ISIS 141923 had no significant effect.

Plasma human PCSK9 levels exhibited time dependent suppression with effects at 6 weeks, greater than at 3 wks for all doses of BMS-844421 (FIG. 16). At 3 wks dosing, only the highest dose of BMS-844421 showed significant suppression. As seen in FIG. 17, data for 6 wks treatment with BMS-844421 showed a sigmoidal dose-response curve, with ED50 approximately 15 mg/kg/dose (given twice weekly). Plotting the mRNA and protein data together showed that the two endpoints generally, though imperfectly, reflect each other, with protein showing greater sensitivity to suppression (FIG. 18).

Male transgenic mice responded more sensitively to BMS-844421 than the female transgenic mice in this study (FIG. 19). The data was not the result of different plasma baseline levels of human PCSK9 between males and females, which is human PCSK9=67+/−31 ng/mL in males and 60+/−27 ng/mL in females in the predosing samples. The difference in PCSK9 protein response for males vs. females was less apparent for liver target mRNA (FIG. 20).

Levels of endogenous mouse PCSK9 mRNA in liver were decreased by ISIS 394816 (and the combination of ISIS 394816 with BMS-844421), but were not decreased by BMS-844421 only (FIG. 21).

A key cholesterol regulatory gene, HMGCoA reductase (HMGR), is known to respond to sterol feedback in the liver. An assay of endogenous mouse liver HMGR mRNA revealed 45% suppression of HMGR with BMS-844421 as well as ISIS 394816 (FIG. 22). These findings are consistent with downregulation of PCSK9 promoting higher levels of LDLR, in turn leading to increased cholesterol in the regulatory pool in liver cells, resulting in downregulation of HMGR. Similar, though quantitatively lesser effects, were also seen for the sterol pathway genes LDLR mRNA and for SREBP2 mRNA.

LDLR protein levels were assayed in liver samples from the same study. Total hepatic membranes were prepared, and western blots of SDS-PAGE gels were assayed with anti-LDLR antibodies. As a control, transferrin receptor (TR) protein levels were measured with anti-TR antibodies. Liver samples from the ASO study were pooled from n=3 for the preparation of the membrane fraction. Pairs of pools for each group were assayed. Treatment with BMS-844421, and especially the combination of BMS-844421 with the murine ASO ISIS 394816, led to increases up to >2-fold in levels of LDLR protein, compared to the level of TR protein (FIG. 23 and FIG. 24).

The ELISA assay uses a monoclonal capture antibody and a polyclonal detector antibody developed against human recombinant PCSK9 produced in baculovirus. The recombinant human PCSK9 protein also serves as standard antigen for the ELISA assay. Cynomolgus monkey plasma PCSK9 was readily detectable using this assay. Human PCSK9 protein in plasma of human PCSK9 transgenic mouse lines was specifically detected with no cross-reactivity to murine PCSK9.

Despite these pharmacodynamic responses to BMS-844421, hemizygous transgenic mice did not exhibit changes in plasma LDL-C or total cholesterol following ASO treatment, including BMS-844421 (Table 36). However, normal chow fed mice have low levels of LDL-C (Table 36). It is plausible that these low LDL-C levels are not readily decreased by increases in LDLR protein. Another possible explanation is that the hemizygous expression of human PCSK9, i.e., the gene specifically targeted by BMS-844421, contributes only about ⅓ to ½ of the total murine+human PCSK9 in this hemizygous transgenic mouse model.

TABLE 36 Plasma lipids at 6 wk treatment with ASOs in transgenic mice Total Both Sexes Triglycerides Cholesterol HDL LDL Saline 72 ± 14  108 ± 8.2 57 ± 6.4 14 ± 1 Missense 64 ± 9.2 110 ± 8.6 57 ± 7.1 15 ± 1 BMS-844421 54 ± 8.6 129 ± 7.3 67 ± 4.6 18 ± 2 (30 mg/kg) BMS-844421 54 ± 9.0 126 ± 9.2 65 ± 5.5 18 ± 1 (15 mg/kg) BMS-844421 53 ± 9.4 118 ± 9.3 61 ± 6.9 17 ± 2 (10 mg/kg) BMS-844421 56 ± 9.7 117 ± 8.5 61 ± 6.1 17 ± 1 (5 mg/kg) Mouse ASO 50 ± 7.0 84.2 ± 2.9  45 ± 3.0 11 ± 2 I-394816 (15 mg/kg) BMS-844421 + 55 ± 9.3  92 ± 5.3 48 ± 4.6 12 ± 2 I-394816 (15 mg/kg each) WT Littermates 60 ± 8.1 111.7 ± 7.2   57 ± 5.3 15 ± 1 There were no changes in ALT or AST (not shown), suggesting that no liver abnormalities were present after 6 wks treatment.

18.3. Conclusions

The recently established human genomic PCSK9 transgenic mouse model responded to human PCSK9 ASO treatment with robust downregulation of human PCSK9 at both the mRNA and protein levels in liver. Plasma human PCSK9 was also decreased with the magnitude of decrease somewhat greater than liver mRNA suppression, suggesting that plasma PCSK9 levels are a good readout for suppression of the target mRNA. The ED50 for both endpoints was approximately 15 mg/kg (per dose given twice weekly). The transgenic mice did not exhibit significant changes in plasma LDL-C or total cholesterol following BMS-844421 or ISIS 394816 treatment, nor for the combination of the two ASOs. A trend for increasing liver LDLR protein was observed following treatment with BMS-844421, suggesting a functional effect consistent with the downregulation of PCSK9 mRNA and protein. Consistent with this was the downregulation of the sterol response gene HMGR in liver.

The limited pharmacodynamic responses in this model appear to be related both to the small LDL window in normal chow diet fed mice, and to the co-expression of the human PCSK9 gene with the endogenous murine PCSK9 gene. In the hemizygous mice, human PCSK9 appears to represent only about ⅓ of the total human+murine PCSK9 expressed. Consistent with this, in a previous study the murine specific PCSK9 ASO ISIS 394816 administered to human apoB-CETP double transgenic mice produced up to 90% suppression of liver PCSK9 mRNA and a corresponding decrease of up to 80% in plasma LDL-C levels. 

1. An antisense compound consisting of the nucleotide sequence selected from the group consisting of: SEQ ID NO: 65, SEQ ID NO: 188, SEQ ID NO: 60, SEQ ID NO: 381, SEQ ID NO: 212, SEQ ID NO: 62, SEQ ID NO: 288, SEQ ID NO: 180, and SEQ ID NO:
 8. 2. The compound of claim 1 selected from the group consisting of: SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 381, and SEQ ID NO:
 188. 3. The compound of claim 1, wherein at least one internucleoside linkage is a phosphorothioate internucleoside linkage.
 4. The compound of claim 1, wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.
 5. The compound of claim 1, wherein at least one nucleoside comprises a modified sugar.
 6. The compound of claim 5, wherein the at least one modified sugar is a bicyclic sugar.
 7. The compound of claim 6, wherein each of the at least one bicyclic sugar comprises a 4′-CH(CH3)-O-2′ bridge.
 8. The compound of claim 5, wherein the modified sugar comprises a 2′-O-methoxyethyl moiety.
 9. The compound of claim 1, comprising at least one tetrahydropyran modified nucleoside, wherein a tetrahydropyran ring replaces the furanose ring.
 10. The compound of claim 9, wherein each of the at least one tetra-hydropyran modified nucleoside has the structure:

wherein Bx is an optionally protected heterocyclic base moiety.
 11. The compound of claim 1, wherein at least one nucleoside comprises a modified nucleobase.
 12. The compound of claim 11, wherein the modified nucleobase is a 5-methylcytosine.
 13. The antisense compound of claim 1, wherein the antisense compound is a modified oligonucleotide comprising: a gap segment consisting of linked deoxynucleosides; a 5′ wing segment consisting of linked nucleosides; and a 3′ wing segment consisting of linked nucleosides; wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; and wherein each nucleoside of each wing segment comprises a modified sugar.
 14. The antisense compound of claim 13, wherein the modified oligonucleotide comprises: a gap segment consisting of ten linked deoxynucleosides; a 5′ wing segment consisting of five linked nucleosides; and a 3′ wing segment consisting of five linked nucleosides; wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment, wherein each nucleoside of each wing segment comprises a 2′-O-methoxyethyl sugar; and wherein each internucleoside linkage is a phosphorothioate linkage.
 15. A composition comprising an antisense compound consisting of the nucleotide sequence selected from the group consisting of: SEQ ID NO: 65, SEQ ID NO: 188, SEQ ID NO: 60, SEQ ID NO: 381, SEQ ID NO: 212, SEQ ID NO: 62, SEQ ID NO: 288, SEQ ID NO: 180, and SEQ ID NO: 8, and a pharmaceutically acceptable carrier or diluent.
 16. The composition of claim 15, wherein the antisense compound is selected from the group consisting of: SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 381, and SEQ ID NO:
 188. 17. The composition of claim 15, further comprising a pharmaceutical agent for separate, sequential, or simultaneous administration with said antisense compound.
 18. The composition of claim 17, wherein the pharmaceutical agent is a lipid lowering compound.
 19. The composition of claim 18, wherein the lipid lowering compound is a HMG-CoA reductase inhibitor, cholesterol absorption inhibitor, MTP inhibitor, antisense compound targeted to ApoB, or any combination thereof.
 20. The composition of claim 19, wherein the HMG-CoA reductase inhibitor is atorvastatin, rosuvastatin, or simvastatin.
 21. The composition of claim 19, wherein the cholesterol absorption inhibitor is ezetimibe.
 22. The composition of claim 19, wherein the HMG-CoA reductase inhibitor is simvastatin, and the cholesterol absorption inhibitor is ezetimibe.
 23. The composition of claim 15, wherein at least one internucleoside linkage in the antisense compound is a phosphorothioate internucleoside linkage.
 24. The composition of claim 15, wherein each internucleoside linkage in the antisense compound is a phosphorothioate internucleoside linkage.
 25. The composition of claim 15, wherein at least one nucleoside in the antisense compound comprises a modified sugar.
 26. The composition of claim 25, wherein the at least one modified sugar is a bicyclic sugar.
 27. The composition of claim 26, wherein each of the at least one bicyclic sugar comprises a 4′-CH(CH3)-O-2′ bridge.
 28. The composition of claim 25, wherein the modified sugar comprises a 2′-O-methoxyethyl moiety.
 29. The composition of claim 15, wherein the antisense compound comprises at least one tetrahydropyran modified nucleoside, wherein a tetrahydropyran ring replaces the furanose ring.
 30. The composition of claim 29, wherein each of the at least one tetrahydropyran modified nucleoside has the structure:

wherein Bx is an optionally protected heterocyclic base moiety.
 31. The composition of claim 15, wherein at least one nucleoside in the antisense compound comprises a modified nucleobase.
 32. The composition of claim 31, wherein the modified nucleobase is a 5-methylcytosine.
 33. The composition of claim 15, wherein the antisense compound is a modified oligonucleotide comprising: a gap segment consisting of linked deoxynucleosides; a 5′ wing segment consisting of linked nucleosides; and a 3′ wing segment consisting of linked nucleosides; wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; and wherein each nucleoside of each wing segment comprises a modified sugar.
 34. The composition of claim 33, wherein the modified oligonucleotide comprises: a gap segment consisting of ten linked deoxynucleosides; a 5′ wing segment consisting of five linked nucleosides; and a 3′ wing segment consisting of five linked nucleosides; wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment, wherein each nucleoside of each wing segment comprises a 2′-O-methoxyethyl sugar; and wherein each internucleoside linkage is a phosphorothioate linkage.
 35. A method comprising administering to an animal an antisense compound consisting of the nucleotide sequence selected from the group consisting of: SEQ ID NO: 65, SEQ ID NO: 188, SEQ ID NO: 60, SEQ ID NO: 381, SEQ ID NO: 212, SEQ ID NO: 62, SEQ ID NO: 288, SEQ ID NO: 180, and SEQ ID NO:
 8. 36. The method of claim 35, wherein the antisense compound is selected from the group consisting of: SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 381, and SEQ ID NO:
 188. 37. The method of claim 35, wherein the animal is a human.
 38. The method of claim 35, wherein administering the compound slows progression and/or ameliorates hypercholesterolemia, acute coronary syndrome, polygenic hypercholesterolemia, mixed dyslipidemia, coronary heart disease, early onset coronary heart disease, type II diabetes, type II diabetes with dyslipidemia, hepatic steatosis, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, hypertriglyceridemia, hyperfattyacidemia, hyperlipidemia, metabolic syndrome, atherosclerosis, or improves cardiovascular outcome, or any combination thereof.
 39. The method of claim 35 further comprising administering at least one pharmaceutical agent.
 40. The method of claim 39, wherein the pharmaceutical agent is administered separately, sequentially, or simultaneously with the antisense compound.
 41. The method of claim 39, wherein the compound and the pharmaceutical agent are administered in the same or separate formulation.
 42. The method of claim 39, wherein the pharmaceutical agent is a lipid lowering pharmaceutical agent.
 43. The method of claim 42, wherein the lipid lowering pharmaceutical agent is a HMG-CoA reductase inhibitor, cholesterol absorption inhibitor, MTP inhibitor, or antisense compound targeted to ApoB, or any combination thereof.
 44. The method of claim 43, wherein the HMG-CoA reductase inhibitor is atorvastatin, rosuvastatin, or simvastatin.
 45. The method of claim 43, wherein the cholesterol absorption inhibitor is ezetimibe.
 46. The method of claim 43, wherein the HMG-CoA reductase inhibitor is simvastatin, and the cholesterol absorption inhibitor is ezetimibe.
 47. The method of claim 35, wherein the administering is parenteral administration.
 48. The method of claim 47, wherein the parenteral administration comprises subcutaneous or intravenous administration.
 49. The method of claim 35, wherein the administering comprises oral administration.
 50. The method of claim 35, wherein the animal has an elevated LDL-cholesterol level above a target level of at least about 100 mg/dL, 130 mg/dL, 160 mg/dL, or 190 mg/dL.
 51. The method of claim 35, wherein the administering the compound to the animal reduces LDL-cholesterol levels.
 52. The method of claim 51, wherein administering the compound reduces the LDL-cholesterol level below a target level of at least about 190 mg/dL, 160 mg/dL, 130 mg/dL, 100 mg/dL, 70 mg/dL, or 50 mg/dL.
 53. The method of claim 35, wherein administering the compound results in reduced coronary heart disease risk, or slows, stops or ameliorates progression of atherosclerosis, or any combination thereof.
 54. The method of claim 37, wherein administering the compound results in improved cardiovascular outcome of the human.
 55. The method of claim 54, wherein the improved cardiovascular outcome is improved carotid intimal media thickness, improved atheroma thickness, increased HDL-cholesterol, or any combination thereof.
 56. The method of claim 35, wherein administering the compound ameliorates hepatic steatosis, results in lipid lowering, improves LDL/HDL ratio, or any combination thereof.
 57. The method of claim 35, wherein at least one internucleoside linkage in the antisense compound is a phosphorothioate internucleoside linkage.
 58. The method of claim 57, wherein each internucleoside linkage in the antisense compound is a phosphorothioate internucleoside linkage.
 59. The method of claim 35, wherein at least one nucleoside in the antisense compound comprises a modified sugar.
 60. The method of claim 59, wherein the at least one modified sugar is a bicyclic sugar.
 61. The method of claim 60, wherein each of the at least one bicyclic sugar comprises a 4′-CH(CH3)-O-2′ bridge.
 62. The method of claim 59, wherein the modified sugar comprises a 2′-O-methoxyethyl moiety.
 63. The method of claim 35, wherein the antisense compound comprises at least one tetrahydropyran modified nucleoside, wherein a tetrahydropyran ring replaces the furanose ring.
 64. The method of claim 63, wherein each of the at least one tetra-hydropyran modified nucleoside has the structure:

wherein Bx is an optionally protected heterocyclic base moiety.
 65. The method of claim 35, wherein at least one nucleoside in the antisense compound comprises a modified nucleobase.
 66. The method of claim 65, wherein the modified nucleobase is a 5-methylcytosine.
 67. The method of claim 35, wherein the antisense compound is a modified oligonucleotide comprising: a gap segment consisting of linked deoxynucleosides; a 5′ wing segment consisting of linked nucleosides; and a 3′ wing segment consisting of linked nucleosides; wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; and wherein each nucleoside of each wing segment comprises a modified sugar.
 68. The method of claim 67, wherein the modified oligonucleotide comprises: a gap segment consisting of ten linked deoxynucleosides; a 5′ wing segment consisting of five linked nucleosides; and a 3′ wing segment consisting of five linked nucleosides; wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment, wherein each nucleoside of each wing segment comprises a 2′-O-methoxyethyl sugar; and wherein each internucleoside linkage is a phosphorothioate linkage. 